Cephalosporin antibiotics

ABSTRACT

Novel cephalosporin antibiotic derivatives.

FIELD OF INVENTION

This invention relates to novel cephalosporin derivatives useful asantibiotics and processes for their preparation.

SUMMARY OF INVENTION

Compounds of the following general Formula I are useful as antibioticagents: ##STR1## wherein Aryl is selected from phenyl and 2-thienyl; Yis selected from hydrogen, chlorine, bromine, a straight or branchedlower alkyl group of from 1 to 4 carbon atoms, and a lower alkoxy groupof from 1 to 4 carbon atoms with the proviso that when Aryl is2-thienyl, Y is hydrogen; Z is selected from a bond, oxygen, sulfur andimino with the proviso that when Aryl is 2-thienyl, Z is a bond; W isselected from hydrogen, methyl, amino, hydroxy, SO₃ H and COOR₁ whereinR₁ is selected from hydrogen and 5-indanyl; n is zero, 1 or 2 with theproviso that when W is other than hydrogen or methyl, and Z is otherthan a bond, n is not zero; R₂ is selected from hydrogen or methoxy; Mis selected from an anion; hydrogen; alkanoyloxymethyl wherein thealkanoyl moiety contains from 1 to 5 carbon atoms and may be straight orbranched; alkanoylaminomethyl wherein the alkanoyl moiety contains from1 to 5 carbon atoms and may be straight or branched and wherein theamino nitrogen may be substituted with an alkyl group of from 1 to 4carbon atoms; alkoxycarbonylaminomethyl wherein the alkoxy moietycontains from 1 to 4 carbon atoms and may be straight or branched andwherein the amino nitrogen atom may be substituted with an alkyl groupof from 1 to 4 carbon atoms; p-(alkanoyloxy)benzyl wherein the alkanoylmoiety contains from 1 to 5 carbon atoms and may be straight orbranched; or aminoalkanoyloxymethyl wherein the alkanoyl moiety containsfrom 2 to 15 carbon atoms, and the amino nitrogen atom may be mono- ordi-substituted with a lower alkyl group of from 1 to 4 carbon atoms;with the proviso that when M is other than an anion, the compounds existin the form of a salt of a pharmaceutically acceptable inorganic ororganic acid; X is selected from hydrogen, acetoxy,1,3,4-thiadiazol-5-ylthio, 3-methyl-1,2,4-thiadiazol-5-ylthio, tetrazol-5-ylthio, 1-methyl-tetrazol-5-ylthio, 2-methyl-1,3,4-oxadiazol-5-ylthio,2-methyl-1,3,4-thiadiazol-5-ylthio and 1,2,3-triazol-5-yl-thio; andpharmaceutically acceptable salts and individual optical isomersthereof. Compounds wherein M is an anion are also known as inner saltsor zwitterions.

DETAILED DESCRIPTION OF INVENTION

In general Formula I the substituent group is represented by M inaddition to being hydrogen or an anion may also be alkanoyloxymethyl asrepresented by the structure: ##STR2## wherein R₃ is selected from astraight or branched lower alkyl group of from 1 to 4 carbon atoms;alkanoylaminomethyl or alkoxycarbonylaminomethyl as represented by thestructure: ##STR3## wherein R₄ represents a straight or branched loweralkyl group of from 1 to 4 carbon atoms or a straight or branched alkoxygroup of from 1 to 4 carbon atoms, and R₅ is selected from hydrogen anda lower alkyl group of from 1 to 4 carbon atoms; p-(alkanoyloxy)benzylas represented by the structure: ##STR4## wherein R₆ is a straight orbranched lower alkyl of from 1 to 4 carbon atoms; andaminoalkanoyloxymethyl as represented by the group: ##STR5## wherein mis 0 to 5, each of R₇ and R₈ is selected from hydrogen or lower alkyl offrom 1 to 4 carbon atoms, and each of R₉ and R₁₀ is selected fromhydrogen or a straight or branched lower alkyl group of from 1 to 4carbon atoms.

Illustrative examples of straight or branched lower alkyl groups of from1 to 4 carbon atoms which Y, R₃, R₄, R₆, R₉ and R₁₀ may represent aremethyl, ethyl, n-propyl, isopropyl, n-butyl and tert-butyl.

Examples of lower alkyl groups of from 1 to 4 carbon atoms which R₅, R₇,and R₈ may represent are methyl, ethyl, n-propyl and n-butyl.

Examples of lower alkoxy groups which Y may represent are methoxy,ethoxy, n-propoxy and n-butoxy.

In general Formula I, the substituent group X may represent in additionto hydrogen or acetoxy, a heterocyclicthio group selected from1,3,4-thiadiazol-5-ylthio,3-methyl-1,2,4-thiadiazol-5-ylthio,tetrazol-5-ylthio, 1-methyltetrazol-5-ylthio,2-methyl-1,3,4-oxadiazol-5-ylthio, 2-methyl-1,3,4-thiadiazol-5-ylthio,or 1,2,3-triazol-5-ylthio as represented by the following respectivestructures: ##STR6##

When the Aryl group in the compounds of general Formula I representphenyl, each of the pyridiniummethyl substituent and the Y substituentmay be individually attached to any of the positions 2 through 6 of thephenyl ring. Compounds of this type may be represented by the followinggeneral Formula II. ##STR7##

In the above Formula II, the substituent groups Y, Z, n, W, R₂, M and Xhave the meanings defined in general Formula I.

The preferred positions of the attachment of the pyridiniummethylsubstituent in the above Formula II are the ortho- and para-positions ofthe phenyl ring.

When the Aryl group in the compounds of general Formula I represents2-thienyl, Y is hydrogen and Z is a bond. Compounds of this type may berepresented by the following Formula III. ##STR8## In the compounds ofthe above Formula III, the pyridiniummethyl substituent may be attachedat the 4 or 5 positions of the thienyl group. In the above Formula III,the substituents as represented by n, W, R₂, M and X have the meaningsdefined in general Formula I.

In the compounds of general Formulas I to III, it is apparent that theR₂ substituent may be either cis or trans to the hydrogen atom at the 6-position of the cephalosporin derivatives. The compounds of Formulas Ito III wherein the R₂ substituent is cis to the aforementioned hydrogenatom are preferred.

Other preferred embodiments of this invention are:

(A) compounds wherein W represents hydrogen, hydroxy, amino, SO₃ H andCOOR₁ wherein R₁ represents hydrogen in that such substitution resultsin compounds having broader spectrum activity and/or improved oralactivity for example wherein:

(1) W represents hydroxy are more resistant to β-lactamase organisms;

(2) W represents SO₃ H or COOR₁ wherein R₁ represents hydrogen havebroader gram negative spectrum;

(3) W represents NH₂ have improved oral activities;

(B) compounds wherein R₂ represents methoxy are of particular interestin that such compounds demonstrate antibacterial activity againstcephalosporinase producing gram negative organisms.

(C) compounds wherein X represents acetoxy,2-methyl-1,3,4-thiadiazol-5-ylthio, or 1-methyltetrazol-5-ylthio. Of thepreferred embodiments set forth in (A), (B) and (C) compounds wherein Zrepresents a bond are more preferred. The most preferred compounds ofthis invention are those represented by the following Formula IV:##STR9## wherein W' is selected from hydrogen, hydroxy, amino, COOH orSO₃ H; X' is selected from hydrogen, acetoxy,2-methyl-1,3,4-thiadiazol-5-ylthio or 1-methyltetrazol-5-ylthio; andpharmaceutically acceptable salts thereof.

In the above Formula IV, compounds wherein the hydrogen atoms at the 6-and 7-positions are cis to one another are preferred.

The individual optical isomers of the compounds of this inventionwherein W or W' is other than hydrogen are also included within thescope of this invention.

The non-toxic inorganic acid addition salts of the compounds of thisinvention such as mineral acid addition salts, for example, hydrogenchloride, hydrogen bromide, hydrogen iodide, sulfates, sulfamate,phosphate, and organic acid addition salts, for example, maleate,acetate, citrate, oxalate, succinate, benzoate, tartrate, fumarate,malate, mandalate, and ascorbate, are also included within the scope ofthis invention.

Also within the scope of this invention are the nontoxicpharmaceutically acceptable salts of the compounds of this inventionwherein W represents COOH or SO₃ H and compounds wherein M representshydrogen. Illustrative pharmaceutically acceptable salts of these acidsderivatives are primary, secondary, or tertiary amines, for example,cyclohexylamine, ethylamine, and pyridine.

The pharmaceutically acceptable cations which may be present as thegroup M in the compounds of general Formulas I to III include alkalimetal ion, for example, sodium ion, potassium ion, calcium ion as wellas ammonium, an organic amine cation, for example, lower alkyl ammoniumgroups, such as triethylammonium, and N-ethyl piperidine.

The salt forms of compounds of Formulas I to III wherein M is apharmaceutically acceptable cation are prepared in the manner recognizedin the art and may be found in situ or by reacting the correspondingacid with base, for example, sodium bicarbonate or triethylamine.

The compounds of this invention may be administered in a manner similarto that of many well-known cephalosporin compounds, for example,cephalexin, cephalothin, or cephaloglycine. They may be administeredalone or in the form of pharmaceutical preparations either orally orparenterally and topically to warm blooded animals, that is, birds, andmammals, for example, cats, dogs, cows, sheep and horses, and humans.For oral administration, the compounds may be administered in the formof tablets, capsules or pills or in the form of elixirs or suspensions.For parenteral administration, they may best be used in the form of asterile aqueous solution which may contain other solutes, for example,enough saline or glucose to make the solution isotonic. For topicaladministration, the compounds may be incorporated in creams orointments.

Illustrative examples of bacteria against which the compounds of thisinvention are active are Staphylococcus aureus, Salmonellaschottmuelleri, Klebsiella pneumoniae, Diplococcus pneumoniae, andStreptococcus pyogenes.

An illustrative example of a cephalosporin derivative of this inventionis3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylicacid. Additional examples of compounds of this invention are set forthhereinbelow in the specific examples which are representative of theinvention.

The compounds of this invention wherein R₁ is hydrogen are prepared bycoupling 7-aminocephalosporanic acid or a derivative thereof having theformula: ##STR10## wherein R₂, M and X have the meaning defined ingeneral Formula I with an acid of the following Formula VI or afunctional derivative thereof: ##STR11## wherein Aryl, Y, Z and n havethe meanings defined in general Formula I, and W₂ is selected fromhydrogen, methyl, amino, hydroxy, SO₃ H, or COOH. When the substituentgroup W₂ in the above Formula VI represents an amino group, suitableblocking groups, for example, an acid salt such as hydrochloride salt,an acyl group, or tert-butoxycarbonyl may be employed to protect theamino function. Such blocking groups are removed after the couplingreaction by the methods generally known in the art, for example, asdescribed by Lemiuex et al., in U.S. Pat. No. 3,657,232.

Functional equivalents of the acids as represented by Formula VI includethe acid halide, for example, the acid chloride, acid anhydrides,including mixed anhydrides with, for example, alkylphosphoric acids,lower aliphatic mono esters of carbonic acid, or alkyl or aryl sulphonicacids. Additionally, the acid azide or an active ester or thioester forexample, with p-nitrophenol, 2,4-dinitrophenol, or thioacetic acid, maybe used or the free acid as represented by Formula VI may be coupledwith the 7-aminocephalosporanic acid derivative as represented byFormula V after first reacting the acid withN,N'-dimethylchloroforminium chloride or by use of a carbodiimidereagent, for example, N,N'-diisopropylcarbodiimide,N,N'-dicyclohexylcarbodiimide, orN-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide. Just as the finalproducts of this invention must exist in a form to satisfy the positivecharge on the pyridinium group so must the compounds of Formula VI, forexample, in the form of a salt of a pharmaceutically acceptableinorganic or organic acid.

The coupling reaction is generally carried out in the presence of asolvent. Suitable solvents include ethyl acetate, acetone, dioxane,acetonitrile, chloroform, ethylene chloride, tetrahydrofuran anddimethylformamide. As hydrophilic solvents are employed, mixtures ofthese solvents with water are also suitable for the above reactions. Thecoupling reaction is generally carried out in the presence of a base,for example, an alkaline bicarbonate. The temperature of the reactionmay vary from -10° to 100° C., and the reaction time may vary from about1/2 hour to 10 hours. The cephalosporin products are isolated byconventional methods.

Compounds of Formula V wherein R₂ is hydrogen, M is hydrogen, or ananion and X is hydrogen or acetoxy are commercially available or may beprepared by the methods well-known in the art. The correspondingcompounds wherein R₂ is methoxy may be prepared by the generalprocedures described in U.S. Pat. No. 3,778,432.

Compounds of Formula V wherein M is alkanoyloxymethyl may be prepared byreacting the corresponding acid in the form of a salt, such as, analkali metal salt or the triethylammonium salt with a compound offormula: ##STR12## wherein halo is chlorine or bromine, and R₃ is astraight or branched lower alkyl group of from 1 to 4 carbon atoms, bythe general procedure described in U.S. Pat. No. 3,655,658.

Compounds of Formula V wherein M is alkanoylaminomethyl oralkoxycarbonylaminomethyl are prepared by treating the sodium salt ofacid derivatives of Formula V in an organic solvent such asdimethylformamide or hexamethylphosphoramide. at room temperature withan equivalent amount of an alkanoylaminomethyl halide or analkoxycarbonylaminomethyl halide for 1/2 to 3 hours after which themixture is poured into ice water. The resulting precipitated product isisolated by standard procedures.

Compounds of Formula V wherein M is p-(alkanoyloxy)-benzyl are preparedby adding two equivalents of the p-(alkanoyloxy)benzyl alcohol to asuspension of sodium salts of acid derivatives of Formula V anddimethylformamide or hexamethylphosphoramide after which the mixture iscooled to 0° C. 1.2 equivalents of dicyclohexylcarbodiimide anddimethylformamide are added dropwise to the mixture with stirring. Themixture is stirred at 0° C. for 1/2 to 3 hours and then an additional 2to 5 hours at room temperature. The formed dicyclohexylurea is removedby filtration and the filtrate is diluted with chloroform, methylenechloride or ethylacetate, washed with water and dried to give theproduct.

Compounds of Formula V wherein M is aminoalkanoyloxymethyl are preparedby mixing a suspension of the sodium salt of an acid of Formula V and anexcess of an appropriate amine protected aminoalkanoyloxymethyl halidein a solvent such as dimethylformamide, hexamethylphosphoramide ordimethylsulfoxide for 2 to 96 hours. The mixture is then diluted with asolvent such as ethylacetate or methylene chloride washed with water,aqueous base, then water. The organic phase is separated and theprecipitate isolated by conventional means followed by deprotection ofthe amine group to give the product.

Compounds of Formula V wherein X is a heterocyclic thio group asdescribed in Formula I are prepared by dissolving 1 equivalent of theacid in the form of a salt, such as, the sodium salt wherein X isacetoxy in about 500 to 2000 ml of water at a temperature of from 50° to80° C. under a nitrogen atmosphere and subsequently adding 1 equivalentof a base, such as, triethylammonium or sodium bicarbonate and 1 to 3equivalents of an appropriate heterocyclicthiol selected from a compoundhaving the following structure: ##STR13## The reaction mixture isstirred at 50° to 90° C. for about 2 to 6 hours after which the water isevaporated, and the residue is taken up in an organic solvent, such as,methanol, ethanol, or dimethylformamide, and precipitated with anorganic solvent, such as, acetonitrile, acetone, ethylacetate orchloroform.

Compounds of general Formula VI are prepared by treating a compound ofthe formula: ##STR14## wherein halo is chlorine or bromine, and Aryl, Y,Z, n and W₂ have the meanings defined in general Formula VI withpyridine in a solvent, such as, a lower alcohol, for example, methanol,ethanol, isopropyl alcohol or n-butanol or dimethylsulphoxide,dimethylformamide or aqueous mixture of these solvents, for from 1/2hour to 24 hours at a temperature range from 0° to 125° C. The productscan be isolated by conventional procedures. In some instances, it may bemore convenient to convert the acid as represented by Formula VII to thecorresponding methyl ester by, for example, treating the acid withdiazomethane at -10° C. then stirring the mixture for about 10 to 30minutes at room temperature.

When the substituent group W₂ in compounds of general Formula VIIrepresents amino, the amino group is protected by a suitable blockinggroup, for example, tert-butoxycarbonyl prior to the treatment withpyridine. The blocking group may be removed after the coupling reactionby a mild acid hydrolysis or hydrogenolysis by procedures known in theart.

The compounds of Formula VII are prepared by direct halomethylation asdescribed hereinbelow of an acid of the formula: ##STR15## wherein Aryl,Y, Z, n and W₂ have the meanings defined in general Formula VI which arecommercially available or are obtained by methods well-known in the art.

When the substituent group W₂ in the compound of Formula VIII representsamino, the amino group is protected by a suitable blocking group as forexample described hereinabove in reference to compounds of generalFormula VII.

The halomethylated derivatives of the compounds of Formula VIII areobtained by several methods. For example, the compound of Formula VIIIwith a source of formaldehyde such as paraformaldehyde, ClCH₂ OCH₃, orformaline solution in the presence of a Lewis acid, such as ZnCl₂,AlCl₃, SnCl₄ or ClSO₃ H in a solvent, such as petroleum ether,chloroform, carbontetrachloride or benzene, at a temperature rangingfrom -10° to 100° C. during which time hydrogen chloride gas or hydrogenbromide gas is bubbled into the reaction mixture will give compounds ofFormula VII.

Additionally, upon reaction of an acid of Formula VIII with trioxane inacetic acid or phosphoric acid at temperatures of from -10° to 100° C.during which time hydrogen bromide or hydrogen chloride gas is bubbledthrough the reaction mixture, compounds of general Formula VII areobtained. Or, the reaction of an acid of Formula VIII in the presence ofa Lewis acid, such as those described hereinabove, with chloromethylether at temperatures of from -10° to 100° C. or the reaction of theacid in acetic acid or concentrated sulfuric acid with dichloromethylether in the presence of zinc chloride will give the compounds ofgeneral Formula VII.

The compounds of Formula VII wherein W₂ represents COOH, and Aryl isphenyl are preferably obtained by treating the corresponding diethylester of Formula VIII with 40% formaline in the presence of anhydrouszinc chloride in benzene at about 50° C. during which time hydrogenchloride or hydrogen bromide gas is bubbled into the reaction mixturefollowed by acid hydrolysis.

Compounds of Formula VII wherein W₂ represents SO₃ H may be obtained bythe halomethylation reactions described using an acid of Formula VIIIwherein W₂ represent SO₃ H or the carboxymethyl ester thereof in whichlatter case the resulting halomethylated compound is converted to thefree COOH by acid hydrolysis.

In the halomethylation of compounds of Formula VIII wherein W₂represents OH it may be advantageous to protect the OH group prior tohalomethylation as described by B. Reichert, et al., Pharmazie vol. 5,10 (1950).

Compounds of this invention wherein R₁ is 5-indanyl are prepared byreacting the corresponding acid, that is, compounds of general Formula Iwherein R₁ is hydrogen with 5-indanol in an inert solvent in thepresence of N,N'-dicyclohexylcarbodiimide at pH of about 2.5 and atemperature of about 20° to 30° C. Equimolar amounts of the reactantsare employed or a slight excess of the 5-indanyl may be used. The molaramount of N,N-dicyclohexylcarbodiimide employed is equivalent to themolar amount of 5-indanol. Suitable solvents for the reaction aredioxane, tetrahydrofuran, ethylacetate, dimethylformamide and methylenechloride.

The compounds of this invention may also be prepared by linking amodified polystyrene containing nitrophenol or hydroxysuccinimide groupswith an acid of general Formula VI by the general procedure described inCanadian patent No. 892,580 issued Feb. 8, 1972, by substituting acompound of general Formula V for the penicillanic acid derivativesdescribed therein.

Compounds of this invention wherein R₁ is hydrogen may also be preparedby solvolysis of a compound of the formula: ##STR16## wherein Aryl, Y,Z, n, R₂, M and X have the meanings defined in general Formula I; halois chlorine or bromine; and W² is hydrogen, methyl, amino, hydroxy, SO₃H or COOH; with pyridine in a solvent, such as, a lower alcohol, forexample, methanol, ethanol, isopropyl alcohol, n-butanol ordimethylsulfoxide, dimethylformamide or aqueous mixtures of thesesolvents. The reaction is carried out from 1/2 hour to 24 hours at atemperature of from 0° to 125° C. The products are isolated byconventional means.

Additionally, the compounds of this invention wherein X represents aheterocyclic thio group selected from 1,3,4-thiadiazol-5-ylthio,3-methyl-1,2,3-thiadiazol-5-ylthio, tetrazol-5-ylthio,1-methyltetrazol-5-ylthio, 2-methyl-1,3,4-oxadiazol-5-ylthio,2-methyl-1,3,4-thiadiazol-5-ylthio or 1,2,3-triazol-5-ylthio and Mrepresents hydrogen or an anion may be prepared by reacting the3-[(acetyloxy)methyl] derivative with the appropriate heterocyclic thiolgroup as represented by the following: ##STR17##

In the above Formulas X and XI the substituent groups Aryl, Y, Z, n, Wand R₂ have the meanings defined in general Formula I, and the moietyS-hetero is selected from 1,3,4-thiadiazol-5-ylthio,3-methyl-1,2,4-thiadiazol-5-ylthio, tetrazol-5-ylthio,1-methyltetrazol-5-ylthio, 2-methyl-1,3,4-oxadiazol-5-ylthio,2-methyl-1,3,4-thiadiazol-5-ylthio or 1,2,3-triazol-5-ylthio. In theabove reaction when M is hydrogen resulting in a carboxy group at the4-position of the cephalosporin ring the compounds exist in the form ofa salt of a pharmaceutically acceptable inorganic or organic acid.

In the above reaction, one equivalent of an aqueous solution of azwitterion at a pH range of 3.5 to 7.5, preferably 5.5 to 6.5, may bereacted with one equivalent of the heterothiol derivative to givecompounds of Formula XI. This reaction can be carried out in about 2 to6 hours at a temperature of from 25° to 90° C.

Compounds of this invention wherein M represents alkanoylaminomethyl oralkoxycarbonylaminomethyl, and W is other than COOH may also be preparedby reacting the corresponding compound wherein M is an anion with 1.5 to2.5 equivalents of an appropriate alkanoylaminomethyl halide oralkoxycarbonylaminomethyl halide each of which may be represented by thestructure: ##STR18## wherein halo is selected from a reactive halogenatom such as chlorine or bromine, R₄ is selected from a straight orbranched lower alkyl group of from 1 to 4 carbon atoms or a straight orbranched lower alkoxy group of from 1 to 4 carbon atoms, and R₅ ishydrogen or a lower alkyl group of from 1 to 4 carbon atoms. Thereactants are stirred from about 1 to 5 hours in dimethylformamide,hexamethylphosphoramide or a similar solvent at a temperature rangingfrom 10° to 45° C. after which the reaction mixture is poured into icewater and decanted. The oily residue is taken up in an organic solventsuch as ethylacetate, methylene chloride or benzene washed with base andthen with water and dried over magnesium sulfate. The organic solutionis evaporated to dryness in vacuo to give the desired ester.

Prior to the above esterification reaction, compounds wherein Wrepresents amino are protected with blocking groups for exampletert-butoxycarbonyl or carbobenzyloxy, such groups being removed oncompletion of the esterification procedure by methods generally known inthe art, for example, by methods set forth in the aforementioned U.S.Pat. No. 3,657,232.

Compounds of this invention wherein M represents p-(alkanoyloxy)benzyland W is other than COOH may also be prepared by reacting molarequivalents of the corresponding acid and a p-(alkanoyloxy)benzylalcohol wherein the alkanoyl moiety contains from 1 to 4 carbon atomsand may be straight or branched. The reactants are dissolved in anorganic solvent such as dimethylformamide or hexamethylphosphoramide andcooled to a temperature of from -15° to 25° C. after which an equivalentquantity of dicyclohexylcarbodiimide in dimethylformamide orhexamethylphosphoramide is added dropwise to the reaction mixture withstirring. Stirring is continued for 1/2 to 2 hours at temperatures offrom -15° to 25° C. and then 4 to 6 hours at from 25° to 45° C. Theformed dicyclohexylurea is removed by filtration, and the filtrate isdiluted with chloroform, ethylacetate or methylene chloride and washedwith water. The organic layer is dried and evaporated to give theproduct.

Compounds of this invention wherein M is alkanoyloxymethyl, and W isother than COOH may also be prepared by reacting the correspondingcompound wherein M is an anion with a compound of the formula: ##STR19##wherein halo is chlorine or bromine and R₃ is a straight or branchedlower alkyl group of from 1 to 4 carbon atoms by the general proceduredescribed in U.S. Pat. No. 3,655,658.

Compounds of this invention wherein M is aminoalkanoyloxymethyl, and Wis other than COOH may be prepared by mixing a suspension of thecorresponding compound wherein M is an anion and an excess of anappropriate amine protected aminoalkanoyloxymethyl halide in a solventsuch as dimethylformamide, hexamethylphosphoramide or dimethylsulfoxidefor 2 to 96 hours. The mixture is then diluted with a solvent such asethylacetate or methylene chloride, washed with water, aqueous base,then water. The organic phase is separated and the precipitate isolatedby conventional means followed by deprotection of the amine group togive the product.

The following specific examples are illustrative of the compounds of theinvention and methods of preparation of the compounds.

EXAMPLE 1 p-Chloromethylphenylacetyl chloride

(A) At a temperature of from -10° to 0° C. hydrogen chloride gas isbubbled through a stirred mixture of 102 g of phenylacetic acid, 67.5 gof paraformaldehyde and 67.5 g of zinc chloride in 1000 ml of petroleumether for one hour. Stirring is continued for about one hour at roomtemperature after which the mixture is refluxed for about 2 hours duringwhich time hydrogen chloride gas is bubbled into the mixture. To thereaction mixture is added 1000 ml each of methylene chloride and water.The organic phase is separated and the aqueous phase is extracted twicewith methylene chloride. The combined organic phases are extracted fourtimes with a saturated sodium bicarbonate solution. The organic neutralphase is dried over anhydrous sodium sulfate, filtered and the solventis removed under vacuum to give a neutral by-product which is furtheridentified in Example 5 below. The basic aqueous phase is separated andacidified with cold concentrated hydrochloric acid to pH 2-3, thenextracted three times with methylene chloride. The methylene chloridefraction is dried over anhydrous sodium sulfate, filtered and thesolventevaporated. The resulting oily acidic product is chromatographedonsilica gel using benzene and benzene-acetone as the eluant to givep-chloromethylphenylacetic acid which is recrystallized from hotchloroform. M.P. 147°-149° C.

(B) A mixture of 1 g of p-chloromethylphenylacetic acid and 6 ml ofthionylchloride is stirred at room temperature for 25 hours after whichthe excessthionyl chloride is removed under vacuum to yieldp-chloromethylphenylacetyl chloride.

When in Example 1 (A) an acid selected from Table I is substituted forphenylacetic acid the respective chloromethyl derivative listed in TableIis obtained which can be converted to the acid chloride by theprocedure ofExample 1 (B).

                  TABLE I                                                         ______________________________________                                        Acid               Chloromethyl derivative                                    ______________________________________                                        hydrotropic acid   p-chloromethylhydrotro-                                                       pic acid                                                   mandelic acid      p-chloromethylmandelic                                                        acid                                                       dihydrocinnamic    p-chloromethyldihydro-                                     acid               cinnamic acid                                              2-methylhydrocin-  p-chloromethyl-2-methyl-                                   namic acid         hydrocinnamic acid                                         3-phenyllactic     3-(p-chloromethylphenyl)-                                  acid               lactic acid                                                4-phenylbutyric    4-(p-chloromethylphenyl)-                                  acid               butyric acid                                               2-methyl-4-phenyl- 2-methyl-4-(p-chloro-                                      butyric acid       methylphenyl)butyric acid                                  2-hydroxy-4-phenyl-                                                                              2-hydroxy-4-(p-chloro-                                     butyric acid       methylphenyl)butyric acid                                  phenoxyacetic acid p-chloromethylphenoxy-                                                        acetic acid                                                2-phenoxypropionic 2-(p-chloromethylphenoxy)-                                 acid               propionic acid                                             4-phenoxybutyric   4-(p-chloromethylphenoxy)-                                 acid               butyric acid                                               2-methyl-4-phenoxy-                                                                              2-methyl-4-(p-chloro-                                      butyric acid       methylphenoxy)butyric                                                         acid                                                       3-phenoxypropionic 3-(p-chloromethylphenoxy)-                                 acid               propionic acid                                             3-phenoxylactic    3-(p-chloromethyl-                                         acid               phenoxy)lactic acid                                        anilinoacetic acid p-chloromethylanilino                                                         acetic acid                                                2-hydroxy-2-(2-thi-                                                                              2-hydroxy-2-[2-(5-chloro-                                  enyl)acetic acid   methyl)thienyl]acetic acid                                 2-anilinopropionic 2-(p-chloromethyl)anilino-                                                    propionic acid                                             4-anilinobutyric   4-(p-chloromethylanilino)-                                 acid               butyric acid                                               3-anilinobutyric   3-(p-chloromethylanilino)-                                 acid               butyric acid                                               phenylthioacetic   p-chloromethylphenyl-                                      acid               thioacetic acid                                            2-phenylthiopro-   2-(p-chloromethylphenyl)-                                  pionic acid        thiopropionic acid                                         4-phenylthiobutyric-                                                                             4-(p-chloromethylphenyl)-                                  acid               thiobutyric acid                                           o-chlorophenyl-    o-chloro-p-chloromethyl-                                   acetic acid        phenylacetic acid                                          ______________________________________                                    

EXAMPLE 2 p-Chloromethylphenylglycine

A mixture of 2.03 g of trifluoroacetylated phenylglycine, 0.8 g of zincchloride in chloromethylether is heated at 65° C. for 12 hours.Theexcess reagent is removed under vacuum, and the residue is dissolvedin CH₂ Cl₂, washed with saturated NaHCO₃ solution then saturated sodiumchloride solution. The neutral organic phase is dried over Na₂ SO₄ andconcentrated to an oil which was purified by column chromatographyyielding the methyl ester of p-chlorophenylglycine which can beconverted to the acid hydrochloride by hydrolysis with aqueoushydrochloric acid. The hydrochloride can be converted to the free baseby adjusting the pH of the aqueous solution to about 5. Similarly, thechloromethyl derivatives listed in Table II may be prepared from thelisted acid.

                  Table II                                                        ______________________________________                                        Acid              Chloromethyl derivative                                     ______________________________________                                        phenylalanine     p-(chloromethylphenyl)-                                                       alanine                                                     2-amino-4-phenyl- 2-amino-4-(p-chloro-                                        butyric acid      methylphenyl)butyric                                                          acid                                                        2-amino-4-phenoxy-                                                                              2-amino-4-(p-chloromethyl-                                  butyric acid      phenoxy)butyric acid                                        3-phenoxyalanine  3-(p-chloromethyl-                                                            phenoxy)alanine                                             2-amino-4-anilino-                                                                              2-amino-4-(p-chloro-                                        butyric acid      methylanilino)butyric                                                         acid                                                        2-amino-4-phenyl- 2-amino-4-(p-chloro-                                        thiobutyric acid  methylphenyl)thio-                                                            butyric acid                                                3-phenylthioalanine                                                                             3-(p-chloromethyl-                                                            phenyl)thioalanine                                          2-(2-thienyl)glycine                                                                            2-[2-(5-chloromethyl)-                                                        thienyl]glycine                                             2-amino-3-(2-thienyl)-                                                                          2-amino-3-[2-(5-chloro-                                     propionic acid    methyl)thienyl]pro-                                                           pionic acid                                                 2-amino-4-(2-thienyl)-                                                                          2-amino-4-[2-(5-chloro-                                     butyric acid      methyl)thienyl]butyric                                                        acid                                                        ______________________________________                                    

EXAMPLE 3 p-Chloromethylphenylmalonic acid

When in the procedure of Example 1 (A) an equivalent amount ofphenylmalonic acid diethyl ester is substituted for phenylacetic acid,p-chloromethylphenylmalonic acid diethyl ester is obtained which yieldsthe corresponding acid upon acid hydrolysis. In a similar manner thechloromethyl derivatives listed in Table III may be prepared when thediethyl ester of the corresponding acid listed in Table III issubstitutedfor phenylmalonic acid diethyl ester.

                  TABLE III                                                       ______________________________________                                        Acid                Chloromethyl derivative                                   ______________________________________                                        2-sulfophenylacetic 2-sulfo-p-chloromethyl-                                   acid                phenylacetic acid                                         3-phenyl-2-sulfopro-                                                                              3-(p-chloromethyl-                                        pionic acid         phenyl)-2-sulfopro-                                                           pionic acid                                               4-phenyl-2-sulfo-   4-(p-chloromethyl-                                        butyric acid        phenyl)-2-sulfobutyric                                                        acid                                                      benzylmalonic acid  p-chloromethylbenzyl-                                                         malonic acid                                              phenethylmalonic    p-chloromethylphen-                                       acid                ethylmalonic acid                                         2-phenoxyethyl-     2-(p-chloromethyl-                                        malonic acid        phenoxy)ethylmalonic                                                          acid                                                      2-phenylthioethyl-  2-(p-chloromethyl-                                        malonic acid        phenyl)thioethyl-                                                             malonic acid                                              anilinomethyl-      p-chloromethylanilino-                                    malonic acid        methylmalonic acid                                        2-thienylmalonic    2-[2-(5-chloromethyl)-                                    acid                thienyl]malonic acid                                      2-thenylmalonic     2-[2-(5-chloromethyl)-                                    acid                thenyl]malonic acid                                       ______________________________________                                    

EXAMPLE 4 5-Chloromethyl-2-thienylacetyl chloride

2-Thiophenecarboxylic acid is treated in a solution of chloroform withchloromethyl ether in the presence of 0.9 to 2.2 equivalents of aluminumchloride to give 5-chloromethyl-2-thienylcarboxylic acid. Treatment oftheobtained acid with excess thionyl chloride at room temperature forabout 16hours yields the acid chloride which is reacted withdiazomethane to give the corresponding diazoketone. A methanol solutionof the diazoketone is irradiated under nitrogen for about one hour witha high pressure mercury lamp using a Quarz filter. The methyl5-chloromethyl-2-thienylacetate is obtained upon work up and columnchromatography on silica gel. The acetateis hydrolyzed by treatment of a1:1 mixture of acetic acid and concentratedhydrochloric acid at roomtemperature overnight to give 5-chloromethyl-2-thienylacetic acid.

When in the procedure of Example 1 (B) 5-chloromethyl-2-thienylaceticacid,is substituted for p-chloromethylphenyl acetic acid,5-chloromethyl-2-thienylacetyl chloride is obtained.

EXAMPLE 5 o-Hydroxymethylphenylacetic acid lactone

The neutral by-product obtained in Example 1 is purified by sublimationunder vacuum (0.05 mm Hg at 80° C.) to give o-hydroxymethylphenylaceticacid lactone. M.P. 82° C.

EXAMPLE 6 o-Bromomethylphenylacetyl chloride

To a solution of 5 ml of glacial acetic acid saturated with hydrogenbromide gas is added at 0° C. a solution of o-hydroxymethylphenylaceticacid lactone (0.55 g) in 2 ml of glacial acetic acid. The mixture isstirred at room temperature for two hours thenrefluxed for one hourduring which time hydrogen bromide gas is bubbled into the mixture. Theexcess lactone and solvent are removed under high vacuum at roomtemperature. The resulting oily residue is triturated threetimes withhexane to give o-bromomethylphenylacetic acid. M.P. 110° C.

A solution of 0.18 g of o-bromomethylphenylacetic acid in excess thionylchloride is stirred at room temperature for 18 hours after which theunreacted thionyl chloride is removed under high vacuum to giveo-bromomethylphenylacetyl chloride as an oily residue.

EXAMPLE 7 o-Chloromethyl-p-methoxymandelic acid chloride

A solution of 1.1 g of 2-chloromethyl-4-methoxymandelic acid, obtainedby the procedure described by B. Reichert et al., Pharmazie 5, 10(1950), in 25 ml of thionyl chloride is stirred at room temperature forabout 16 hours after which the excess thionyl chloride is removed underhigh vacuumto give o-chloromethyl-p-methoxymandelic acid chloride as anoil.

EXAMPLE 83-[(Acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester

To 35 ml of dimethyl formamide is added 7.5 g of the sodium salt of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid, and the solution is stirred at room temperature for about 30minutes after which 8 ml of chloromethylpropionate is added. Stirring iscontinued at room temperature for about 3 hours. The mixture isdilutedwith ethyl acetate and washed with water. The organic layer isseparated and evaporated to dryness. The residue is recrystallized fromethyl acetate to give3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester.

In a similar manner when an appropriate amount of chloromethylpivalate,chloromethylacetate or chloromethylbutyrate is substituted forchloromethylpropionate, the following respective products are obtained:

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid acetyloxymethyl ester,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid butyryloxymethyl ester.

EXAMPLE 93-[(Acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid 2-amino-3-methylbutyryloxymethyl ester

A suspension of 5 grams of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid sodium salt and 8.5 grams ofN-tert-butoxycarbonyl-L-valinechloromethyl ester, which is prepared bythe general procedure described inW. German Offen. No. 2,236,620, aremixed in 100 ml of dimethyl formamide and stirred for 72 hours. Themixture is diluted with ethyl acetate, washed with water with aqueousbicarbonate and again with water. The organic layer is dried overmagnesium sulfate, filtered, and evaporated todryness to give3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-tert-butoxycarbonyl-2-amino-3-methylbutyryloxymethyl ester fromwhich the amine protecting group is removed by standard procedures togive the title product.

EXAMPLE 103-[(Acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-ethoxycarbonyl-N-methylaminomethyl ester

725 mg (2.5 mM) of the sodium salt of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in 50 ml of dimethyl formamide is treated at room temperature with375 mg (2.5 mM) of N-chloromethyl-N-methylurethane for one hour. Themixture is carefully poured into ice water and the precipitated solid isremoved by filtration and washed with water. The solid is dissolved inethylacetate and washed with aqueous sodium bicarbonate and then withwater. The organic layer is dried over magnesiumsulfate filtered andevaporated to dryness in vacuo to give3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-ethoxycarbonyl-N-methylaminomethyl ester.

When in the above procedure an appropriate amount ofN-methyl-N-propionylaminomethyl chloride, N-butyrylaminomethyl chloride,N-acetylaminomethyl chloride, or N-methyl-N-ethoxycarbonylaminomethylchloride is substituted for N-chloromethyl-N-methylurethane thefollowing respective compounds are obtained:

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-methyl-N-propionylaminomethyl ester,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-butyrylaminomethyl ester,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-acetylaminomethyl ester and,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-methyl-N-ethoxy-carbonylaminomethyl ester.

EXAMPLE 113-[(Acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-pivalyloxybenzyl ester

To a suspension of 6.6 mM of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid sodium salt in 35 ml of dimethyl formamide (DMF) is added2equivalents of p-pivalyloxybenzyl alcohol followed by cooling to 0° C.after which 7.2 mM of dicyclohexylcarbodiimide in 7.5 ml of DMF is addeddropwise with stirring. The mixture is stirred at 0° C. for one hour andan additional four hours at room temperature. The formeddicyclohexylurea is removed by filtration. The filtrate is diluted withchloroform, washed with water, dried over magnesium sulfate, filtered,andevaporated in vacuo to give3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-pivalyloxybenzyl ester.

When in the above procedure an appropriate amount ofp-(propionyloxy)benzylalcohol, p-(acetyloxy)benzyl alcohol, orp-(valeryloxy)benzyl alcohol is substituted for p-pivalyloxybenzylalcohol the following respective products are obtained:

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-(propionyloxy)benzyl ester,

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-(acetyloxy)benzyl ester, and

3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-(valeryloxy)benzyl ester.

EXAMPLE 123[(2-Methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

In about 1 liter of water is dissolved 0.1 mole of the sodium salt of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid at 70° C. under nitrogen atmosphere. To the solutionis added 1equivalent of sodium bicarbonate and 2 equivalents of2-methyl-1,3,4-thiadiazol-5-ylthiol. The mixture is stirred at 70° C.for 3 hours after which the pH is adjusted to 3.5, and the resultingprecipitate collected giving3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

When in the above procedure an equivalent amount of1,3,4-thiadiazol-5-ylthiol, 3-methyl-1,2,4-thiadiazol-5-ylthiol,tetrazol-5-ylthiol, 1-methyltetrazol-5-ylthiol or2-methyl-1,3,4-oxadiazol-5-ylthiol is substituted for2-methyl-1,3,4-thiadiazol-5-ylthiol the following respective productsare obtained:

3-[(1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid,

3-[(3-methyl-1,2,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid,

3-[(tetrazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylicacid,

3-[(1-methyltetrazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid, and

3-[(2-methyl-1,3,4-oxadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

EXAMPLE 13

When in the procedure of Example 12 appropriate amounts of the sodiumsalt of the cephalosporin derivative and the heterocyclicthiolderivative listed below in Table IV are substituted respectively for thesodium salt of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and 2 -methyl-1,3,4-thiadiazol-5-ylthiol the respective productslisted in Table IV are obtained.

                                      TABLE IV                                    __________________________________________________________________________    Cephalosporin Derivative                                                                             Heterocyclicthiol                                                                             Product                                __________________________________________________________________________    3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              3-methyl-1,2,4-thiadiazol-5-                                                                  3-[(3-methyl-1,2,4-thiadiazol-5-          5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                              ylthiol         ylthio)methyl]-7-amino-8-oxo-5-thia-      2-carboxylic acid propionyloxymethyl                                                                              1-azabicyclo[4.2.0]oct-2-ene-2-car-       ester                               boxylic acid propionyloxymethyl                                               ester                                     3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              1-methyltetrazol-5-ylthiol                                                                    3-[(1-methyltetrazol-5-ylthio)methyl]-                                        1                                         5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              7-amino-8-oxo-5-thia-1-azabicyclo-        2-carboxylic acid p-pivalyloxymethyl                                                                              [4.2.0]oct-2-ene-2-carboxylic acid        ester                               pivalyloxymethyl ester                    3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              1,3,4-thiadiazol-5-ylthiol                                                                    3-[(1,3,4-thiadiazol-5-ylthio)methyl]-                                        9                                         5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              7-amino-8-oxo-5-thia-1-azabicyclo-        2-carboxylic acid acetyloxymethyl   [4.2.0]oct-2-ene-2-carboxylic acid        ester                               acetyloxymethyl ester                     3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              tetrazol-5-ylthiol                                                                            3-[(tetrazol-5-ylthio)methyl]-7-amino-                                        .                                         5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              8-oxo-5-thia-1-azabicyclo[4.2.0]oct-      2-carboxylic acid butyryloxymethyl  2-ene-2-carboxylic acid butyryloxy-       ester                               methyl ester                              3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              2-methyl-1,3,4-oxadiazol-                                                                     3-[(2-methyl-1,3,4-oxadiazol-5-yl-        5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                              5-ylthio        thio)methyl]-7-amino-8-oxo-5-thia-1-      2-carboxylic acid 2-amino-3-methyl- azabicyclo[4.2.0]oct-2-ene-2-carbox-      butyryloxymethyl ester              ylic acid 2-amino-3-methylbutyryloxy-                                         methyl ester                              3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              2-methyl-1,3,4-thiadiazol-5-                                                                  3-[(2-methyl-1,3,4-thiadiazol-5-yl-       5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                              ylthiol         thio)methyl]-7-amino-8-oxo-5-thia-        2-carboxylic acid N-ethoxycarbonyl- 1-azabicyclo[4.2.0]                                                           oct-2-ene-2-car-                          N-methylaminomethyl ester           boxylic acid N-ethoxycarbonyl-N-                                              methylaminomethyl ester                   3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              1-methyltetrazol-5-ylthiol                                                                    3-[(1-methyltetrazol-5-ylthio)methyl]-    5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              7-amino-8-oxo-5-thia-1-azabicyclo-        2-carboxylic acid p-pivalyloxybenzyl                                                                              [4.2.0]oct-2-ene-2-carboxylic acid        ester                               p-pivalyloxybenzyl ester                  3-[(acetyloxy)methyl]-7-amino-7-                                                                  2-methyl-1,3,4-thiadiazol-                                                                    3-[(2-methyl-1,3,4-thiadiazol-5-yl-       methoxy-8-oxo-5-thia-1-azabicyclo-                                                                5-ylthiol       thio)methyl]-7-amino-7-methoxy-8-oxo-     [4.2.0]oct-2-ene-2-carboxylic acid  5-thia-1-azabicyclo[4.2.0]oct-2-          (prepared by acid hydrolysis of the ene-2-carboxylic acid                     corresponding benzhydryl ester                                                described in U.S. patent 3,778,432)                                           3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              1-methyltetrazol-5-ylthiol                                                                    3-[(1-methyltetrazol-5-ylthio)methyl-     5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              7-amino-8-oxo-5-thia-1-azabicyclo-        2-carboxylic acid p-(propionyloxy)- [4.2.0]oct-2-ene-2-carboxylic acid        benzyl ester                        p-(propionyloxy)benzyl ester              3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              tetrazol-5-ylthiol                                                                            3-[(tetrazol-5-ylthio)methyl-7-amino-     5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                                              8-oxo-5-thia-1-azabicyclo[4.2.0]oct-      2-carboxylic acid p-(acetyloxy)benzyl                                                                             2-ene-2-carboxylic acid                                                       p-(acetyloxy)-                            ester                               benzyl ester                              3-[(acetyloxy)methyl]-7-amino-8-oxo-                                                              3-methyl-1,2,4-thiadiazol-5-                                                                  3-[(3-methyl-1,2,4-thiadiazol-5-          5-thia-1-azabicyclo[4.2.0]oct-2-ene-                                                              ylthiol         ylthio)methyl]-7-amino-8-oxo-5-           2-carboxylic acid p-(valeryloxy)benzyl                                                                            thia-1-azabicyclo[4.2.0]oct-2-            ester                               ene-2-carboxylic acid p-(valery-                                              oxy)benzyl ester                          __________________________________________________________________________

EXAMPLE 143-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

A mixture of 1 g of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and 1 g of p-chloromethylphenylacetyl chloride in 45 ml of ethylacetate is refluxed for about 2 hours after which the solvent is removedunder vacuum yielding a yellow-brown amorphous product which ischromatographed on silica gel using benzene-acetone as the eluant togive3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid. M.P. 164°-165° C. (dec.).

When in the procedure of Example 14 an appropriate amount of an acidchloride listed in the following Table V is substituted forp-chloromethylphenylacetyl chloride the respective cephalosporinderivatives listed in Table V are obtained. The acid chloridederivatives listed in the following Table V are obtained from thecorresponding acid listed in Table I by treatment with thionyl chlorideby the general procedure described in Example 1 (B).

                  TABLE V                                                         ______________________________________                                                          CEPHALOSPORIN                                               ACID CHLORIDE     DERIVATIVE                                                  ______________________________________                                        p-chloromethylhydrotropic acid                                                                  3-[(acetyloxy)methyl]-7-[[2-                                chloride          [4-(chloromethyl)phenyl]-2-                                                   methylacetyl]amino]-8-oxo-                                                    5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 p-chloromethyldihydrocyn-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                namic acid chloride                                                                             [4-(chloromethyl)phenyl]pro-                                                  pionyl]amino]-8-oxo-5-thia-                                                   1-azabicyclo[4.2.0]oct-2-                                                     ene-2-carboxylic acid                                       p-chloromethyl-2-methylhydro-                                                                   3-[(acetyloxy)methyl]-7-[[3-                                cynnamic acid chloride                                                                          [4-(chloromethyl)phenyl]-2-                                                   methylpropionyl]amino]-8-oxo-                                                 5-thia-1-azabicyclo[4.2.0]oct-                                                2-ene-2-carboxylic acid                                     4-(p-chloromethylphenyl)-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                butyric acid chloride                                                                           [4-(chloromethyl)phenyl]bu-                                                   tyryl]amino]-8-oxo-5-thia-1-                                                  azabicyclo[4.2.0]oct-2-ene-2-                                                 carboxylic acid                                             2-methyl-4-(p-chloromethyl-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                phenyl)butyric acid chloride                                                                    [4-(chloromethyl)phenyl]-2-                                                   methylbutyryl]amino]-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 p-chloromethylphenoxyacetic                                                                     3-[(acetyloxy)methyl]-7-[[2-                                acid chloride     [4-(chloromethyl)phenoxy]-                                                    acetyl]amino]-8-oxo-5-thia-1-                                                 azabicyclo[4.2.0]oct-2-ene-2-                                                 carboxylic acid                                             2-(p-chloromethylphenoxy)-                                                                      3-[(acetyloxy)methyl]-7-[[2-                                propionic acid chloride                                                                         [4-(chloromethyl)phenoxy]-2-                                                  methylacetyl]amino]-8-oxo-5-                                                  thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                     4-p-chloromethylphenoxy)bu-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                tyric acid chloride                                                                             [4-(chloromethyl)phenyl]bu-                                                   tyryl]amino]-8-oxo-5-thia-1-                                                  azabicyclo[4.2.0]oct-2-ene-2-                                                 carboxylic acid                                             2-methyl-4-(p-chloromethyl-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                phenoxy)butyric acid chloride                                                                   [4-(chloromethyl)phenoxy]-2-                                                  methylbutyryl]amino[-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 3-(p-chloromethylphenoxy)-                                                                      3-[(acetyloxy)methyl]-7-[[3-                                propionic acid    [4-(chloromethyl)phenoxy]-                                                    propionyl]amino]-8-oxo-5-                                                     thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                     p-chloromethylanilinoacetic                                                                     3-[(acetyloxy)methyl]-7-[[2-                                acid chloride     [4-(chloromethyl)anilino]-                                                    acetyl]amino]-8-oxo-5-thia-                                                   1-azabicyclo[4.2.0]oct-2-ene-                                                 2-carboxylic acid                                           2-(p-chloromethyl)anilino-                                                                      3-[(acetyloxy)methyl]-7-[[2-                                propionic acid chloride                                                                         [4-(chloromethyl)anilino]-2-                                                  methylacetyl]amino-8-oxo-5-                                                   thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                     4-(p-chloromethylanilino)-                                                                      3-[(acetyloxy)methyl]-7-[[4-                                butyric acid chloride                                                                           [4-(chloromethyl)anilino]-                                                    butyryl]amino]-8-oxo-5-thia-                                                  1-azabicyclo[4.2.0]oct-2-ene-                                                 2-carboxylic acid                                           3-(p-chloromethylanilino)-                                                                      3-[(acetyloxy)methyl]-7-[[3-                                butyric acid chloride                                                                           [4-(chloromethyl)anilino]-2-                                                  methylpropionyl]amino]-8-oxo-                                                 5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-carboxylic acid                                   p-chloromethylphenylthio-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                acetic acid chloride                                                                            [4-(chloromethyl)phenylthio]-                                                 acetyl]amino]-8-oxo-5-thia-1-                                                 azabicyclo[4.2.0]oct-2-ene-                                                   2-carboxylic acid                                           2-(p-chloromethylphenyl)-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                thiopropionic acid chloride                                                                     [4-(chloromethyl)phenylthio]-                                                 2-methylacetyl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-carboxylic acid                                   4-(p-chloromethylphenyl)-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                thiobutyric acid chloride                                                                       [4-(chloromethyl)phenylthio]-                                                 butyryl]amino]-8-oxo-5-thia-                                                  1-azabicyclo[4.2.0]oct-2-ene-                                                 2-carboxylic acid                                           o-chloro-p-chloromethyl-                                                                        3-[(acetyloxy)methyl]-7-[ [2-                               phenylacetic acid chloride                                                                      [2-(chloro)-4-(chloromethyl)-                                                 phenyl]acetyl]amino]-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-carboxylic acid                                   5-chloromethyl-2-thienylace-                                                                    3-[(acetyloxy)methyl]-7-[[2-                                tyl chloride      [5-(chloromethyl)-2-thienyl]-                                                 acetyl]amino]-8-oxo-5-thia-1-                                                 azabicyclo[4.2.0]oct-2-ene-                                                   2-carboxylic acid                                           2-chloromethyl-4-methoxyman-                                                                    3-[(acetyloxy)methyl]-7-[[2-                                delic acid chloride                                                                             [2-(chloromethyl)-4-methoxy-                                                  phenyl]-2-hydroxyacetyl]-                                                     amino]-8-oxo-5-thia-1-azabi-                                                  cyclo[4.2.0]oct-2-ene-2-car-                                                  boxylic acid                                                4-chloromethylmandelic                                                                          3-[(acetyloxy)methyl]-7-[[2-                                acid chloride     [4-(chloromethyl)phenyl]-2-                                                   hydroxyacetyl]amino]-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 3-(p-chloromethylphenyl)-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                lactic acid chloride                                                                            [4-(chloromethyl)phenyl]-2-                                                   hydroxypropionyl]amino]-8-                                                    oxo-5-thia-1-azabicyclo[                                                      4.2.0]-oct-                                                                   2-ene-2-carboxylic acid                                     2-hydroxy-4-(p-chloro-                                                                          3-[(acetyloxy)methyl]-7-[[4-                                methylphenyl)butyric acid                                                                       [4-(chloromethyl)phenyl]-2-                                 chloride          hydroxybutyryl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 3-(p-chloromethylphenoxy)-                                                                      3-[(acetyloxy)methyl]-7-[[3-                                lactic acid chloride                                                                            [4-(chloromethyl)phenoxy]-                                                    2-hydroxypropionyl]amino]-8-                                                  oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                                        2-hydroxy-2-[2-(5-chloro-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                methyl)thienyl]acetic acid                                                                      [5-(chloromethyl)-2-thienyl]-                               chloride          2-hydroxyacetyl]amino]-8-oxo-                                                 5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 ______________________________________                                    

EXAMPLE 157-[[2-[4-(Chloromethyl)phenyl]acetyl]amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

By the procedure of Example 14, only substituting 1 g of7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid for3-acetyloxymethyl-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid,7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid is obtained. M.P. 145°-146° C.

EXAMPLE 163-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

p-Chloromethylphenylglycine wherein the amino group is protected withtert-butoxycarbonyl, is treated with isobutyl chloroformate in thepresence of triethylamine. The thus obtained mixed anhydride is reactedwith the triethylamine salt of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid at 0° C. for about 4hours. The resulting product is isolated and the amine protecting groupis removed by acid hydrolysis to give3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

When an appropriate amount of an amino acid listed in the followingTable VI is substituted for p-chloromethylphenylglycine in the aboveExample 16,the corresponding cephalosporin derivative listed in thefollowing Table VIis obtained.

                  TABLE VI                                                        ______________________________________                                                         CEPHALOSPORIN                                                AMINO ACID       DERIVATIVE                                                   ______________________________________                                        3-(p-chloromethylphenyl)-                                                                      3-[(acetyloxy)methyl]-7-[[3-                                 alanine          [4-(chloromethyl)phenyl]-2-                                                   aminopropionyl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  2-amino-4-(p-chloromethyl-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                 phenyl)butyric acid                                                                            [4-(chloromethyl)phenyl]-2-                                                   aminobutyryl]amino]-8-oxo-5-                                                  thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                      2-amino-4-(p-chloromethyl-                                                                     3-](acetyloxy)methyl]-7-[[4-                                 phenoxy)butyric acid                                                                           [4-(Chloromethyl)phenoxy]-                                                    2-aminobutyryl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  3-(p-chloromethylphenoxy)-                                                                     3-[(acetyloxy)methyl]-7-[[3-                                 alanine          [4-(chloromethyl)phenoxy]-2-                                                  aminopropionyl)amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  2-amino-4-(p-chloromethyl-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                 anilino)butyric acid                                                                           [4-(chloromethyl)anilino]-2-                                                  aminobutyryl]-8-oxo-5-thia-1-                                                 azabicyclo[4.2.0]oct-2-ene-                                                   2-carboxylic acid                                            2-amino-4-(p-chloromethyl-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                 phenyl)thiobutyric acid                                                                        [4-(chloromethyl)phenylthio]-                                                 2-aminobutyryl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  3-(p-chloromethylphenyl)-                                                                      3-[(acetyloxy)methyl]-7-[[3-                                 thioalanine      [4-(chloromethyl)phenylthio]-                                                 2-aminopropionyl]amino]-8-oxo-                                                5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  2-[2-(5-chloromethyl)thi-                                                                      3-[(acetyloxy)methyl]-7-[[2-                                 enyl]glycine     [5-(chloromethyl)-2-thienyl]-                                                 2-aminoacetyl]amino]-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  2-amino-3-[2-5-chloro-                                                                         3-[(acetyloxy)methyl]-7-[[3-                                 methyl)thienyl]propionic                                                                       [5-(chloromethyl)-2-thienyl]-                                acid             2-aminopropionyl]amino]-8-                                                    oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                                         2-amino-4-[2-(5-chloro-                                                                        3-[(acetyloxy)methyl]-7-[[4-                                 methyl)thienyl]butyric acid                                                                    [5-(chloromethyl)-2-thienyl]l-                                                2-aminobutyryl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                  ______________________________________                                    

EXAMPLE 173-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

α-Carboxy-p-chloromethylphenylacetyl nitrophenyl polymer, preparedaccording to the procedure described in Canadian Pat. No. 892,580,carrying 4 m. mole of p-chloromethylphenylmalonic acid is stirred forabout 8 hours in 20 ml of dry methylene chloride solution containing 1m. mole of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid triethylammonium salt, which is prepared from 544 mg of7-aminocephalosporanic acid (1 m. mole) and 0.56 ml of triethylamine (1m.mole) at room temperature. After only traces of 7-aminocephalosporanicacidremain in solution, which is determined by thin layer chromatographyon cellulose in 70% aqueous propanol, the polymer is filtered off andwashed with 3 portions of 50 ml each of methylene chloride. The combinedfiltrates are evaporated and the residue is dissolved in 20 ml ofdistilled water. This solution is acidified to pH 2 by adding 0.2Nhydrochloric acid and extracted twice with ethyl acetate. The organicsolution is dried over sodium sulfate and evaporated at roomtemperature. The remaining solid is dried over night over phosphoruspentoxide under vacuum to give 3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

When in the procedure of Example 17 an appropriate amount of an acidlistedin the following Table VII is substituted forp-chloromethylphenylmalonic acid the respective cephalosporin derivativelisted in Table VII are obtained.

                  TABLE VII                                                       ______________________________________                                                          CEPHALOSPORIN                                               ACID              DERIVATIVES                                                 ______________________________________                                        2-sulfo-p-chloromethylphenyl-                                                                   3-[(acetyloxy)methyl]-7-[[2-                                acetic acid       [4-(chloromethyl)phenyl]-2-                                                   sulfoacetyl]amino]-8-oxo-5-                                                   thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                     4-(p-chloromethylphenyl)-2-                                                                     3-[(acetyloxy)methyl]-7-[[4-                                sulfobutyric acid [4-(chloromethyl)phenyl]-2-                                                   sulfobutyryl]amino]-8-oxo-5-                                                  thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                                     p-chloromethylbenzylmalonic                                                                     3-[(acetyloxy)methyl]-7-[[3-                                acid              [4-(chloromethyl)phenyl]-2-                                                   carboxypropionyl]amino]-8-                                                    oxo-5-thia-1-azabicyclo[4.2.0]-                                               oct-2-ene-2-carboxylic acid                                 2-(p-chloromethylphenoxy)-                                                                      3-[(acetyloxy)methyl)-7-[[4-                                ethylmalonic acid [4-(chloromethyl)phenoxy]-2-                                                  carboxybutyryl]amino]-8-oxo-                                                  5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                 2-(p-chloromethylphenyl)-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                thioethylmalonic acid                                                                           [4-(chloromethyl)phenylthio]-                                                 2-carboxybutyryl]amino]-8-                                                    oxo-5-thia-1-azabicylco-                                                      [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                                        p-chloromethylanilinomethyl-                                                                    3-[(acetyloxy)methyl]-7-[[3-                                malonic acid      [4-(chlorometyl)anilino[-2-                                                   carboxypropionyl]amino]-8-                                                    oxo-5-thia-1-azabicyclo[4.2.0]-                                               oct-2-end-2-carboxylic acid                                 2-[2-(5-chloromethyl)-                                                                          3-[(acetyloxy)methyl]-7-[[3-                                thienyl]malonic acid                                                                            [5-(chloromethyl-2-thienyl]-                                                  2-carboxypropionyl]amino]-8-                                                  oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                                        ______________________________________                                    

EXAMPLE 183-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxy methyl ester

A mixture of 1 g of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxy methyl ester and 1 g of p-chloromethylphenyl acetylchloride in 45 ml of ethyl acetate is refluxed for about 2 hours afterwhich the solvent is removed under vacuum yielding a product which ischromatographed on silica gel using benzene-acetone as the eluant togive3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxy methyl ester.

When in the procedure of Example 18 an appropriate amount of a7-aminocephalosporanic acid derivative listed in the following TableVIII is substituted for3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxy methyl ester and an appropriate amount of the acidchloride listed in the following Table VIII is substituted forp-chloromethylphenylacetyl chloride, the respective products listed inTable VIII are obtained.

                                      TABLE VIII                                  __________________________________________________________________________                    7-AMINOCEPHALOSPORANIC                                        ACID CHLORIDE   ACID DERIVATIVE    PRODUCT                                    __________________________________________________________________________    p-chloromethylhydrotropic                                                                     3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   acid chloride   8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid pro-                                                                 phenyl]-2-methylacetyl]-                                   pionyloxymethyl ester                                                                            amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid                                                         propionyloxymethyl ester                   p-chloromethyl-2-methyl-                                                                      3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   hydrocynnamic acid chloride                                                                   8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[3-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid ace-                                                                 phenyl]-2-methylpropionyl]-                                tyloxymethyl ester amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid ace-                                                    tyloxymethyl ester                         2-methyl-4-(p-chloromethyl-                                                                   3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   phenyl)butyric acid chloride                                                                  8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[4-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid                                                                      phenyl]-2-methylbutyryl]-                                  butyryloxymethyl ester                                                                           amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid                                                         butyryloxymethyl ester                     2-(p-chloromethylphenoxy)-                                                                    3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   propionic acid chloride                                                                       8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid 2-                                                                   phenoxy]-2-methylacetyl]-                                  amino-3-methylbutyryloxy-methyl                                                                  amino]-8-oxo-5-thia-1-                                     ester              azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid 2-                                                      amino-3-methylbutyryloxy-                                  methyl ester                                                  2-methyl-4-(p-chloromethyl-                                                                   3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   phenoxy)butyric acid chloride                                                                 8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[4-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid N-                                                                   phenoxy[-2-methylbutyryl]-                                 ethoxycarbonyl-N-methylamino-                                                                    amino]-8-oxo-5-thia-1-                                     methyl ester       azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid N-                                                      ethoxycarbonyl-N-methyl-                                                      aminoethyl ester                           p-chloromethylanilinoacetic                                                                   3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   acid chloride   8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid N-                                                                   anilino]acetyl]amino]-8-                                   methyl-N-propionylaminomethyl                                                                    oxo-5-thia-1-azabicyclo-                                   ester              [4.2.0]oct-2-ene-2-car-                                                       boxylic acid N-methyl-N-                                                      propionylaminomethyl                                                          ester                                      4-(p-chloromethylanilino)-                                                                    3[(acetyloxy)methyl]-7-amino-                                                                    3-[(acetyloxy)methyl]-7-                   butyric acid chloride                                                                         8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[4-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid N-                                                                   anilino]butyryl]amino]-                                    butyrylaminomethyl ester                                                                         8-oxo-5-thia-1-azabi-                                                         cyclo[4.2.0]oct-2-ene-2-                                                      carboxylic acid N-butyryl-                                                    aminomethyl ester                          p-chloromethylphenylthio-                                                                     3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   acetic acid chloride                                                                          8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid N-                                                                   phenylthio]acetyl]amino]-                                  acetylaminomethyl ester                                                                          8-oxo-5-thia-1-azabi-                                                         cyclo[4.2.0]oct-2-ene-2-                                                      carboxylic acid N-acetyl-                                                     aminomethyl ester                          4-(p-chloromethylphenyl-                                                                      3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   thiobutyric acid chloride                                                                     8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[4-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid N-                                                                   phenylthio]butyryl]amino]-                                 methyl-N-ethoxycarbonylaminomethyl                                                               8-oxo-5-thia-1-azabicy-                                    ester              clo[4.2.0]oct-2-ene-2-car-                                                    boxylic acid N-methyl-N-                                                      ethoxycarbonylaminomethyl                                                     ester                                      5-chloromethyl-2-thienyl-                                                                     3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   acetyl chloride 8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[5-(chloromethyl)-2-                                   oct-2-ene-2-carboxylic acid p-                                                                   thienyl]acetyl]amino]-8-                                   pivalyloxybenzyl ester                                                                           oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-car-                                                       boxylic acid p-pivalyloxy-                                                    benzyl ester                               p-chloromethylmandelic                                                                        3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   acid chloride   8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid p-                                                                   phenyl]-2-hydroxyacetyl]-                                  (propionyloxy)benzyl ester                                                                       amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid                                                         p-(propionyloxy)benzyl                                                        ester                                      2-hydroxy-4-(p-chloromethyl-                                                                  3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   phenyl)butyric acid chloride                                                                  8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[4-[4-(chloromethyl)-                                     oct-2-ene-2-carboxylic acid p-                                                                   phenyl]-2-hydroxybutyryl]-                                 (acetyloxy)benzyl ester                                                                          amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid p-                                                      (acetyloxy)benzyl ester                    2'-hydroxy-2'-[2-(5-                                                                          3-[(acetyloxy)methyl]-7-amino-                                                                   3-[(acetyloxy)methyl]-7-                   chloromethyl)thienyl]-                                                                        8-oxo-5-thia-1-azabicyclo[4.2.0]-                                                                [[2-[5-(chloromethyl)-2-                   acetic acid chloride                                                                          oct-2-ene-2-carboxylic acid p-                                                                   thienyl]-2-hydroxyacetyl]-                                 (valyryloxy)benzyl ester                                                                         amino]-8-oxo-5-thia-1-aza-                                                    bicyclo[4.2.0]oct-2-ene-                                                      2-carboxylic acid p-(val-                                                     yryloxy)benzyl ester                       p-chloromethylphenyl                                                                          3-[(2-methyl-1,3,4-thiadiazol-5-                                                                 3-[(2-methyl-1,3,4-thia-                   acetyl chloride ylthio)methyl]-7-amino-8-oxo-5-                                                                  diazol-5-ylthio)methyl]-7-                                 thia-1-azabicyclo[4.2.0]oct-2-ene-                                                               [[2-[4-(chloromethyl)phen-                                 2-carboxylic acid  yl]acetyl]amino]-8-oxo-5-                                                     thia-1-azabicyclo[4.2.0]-                                                     oct-2-ene-2-carboxylic                                                        acid                                       p-chloromethylphenoxy-                                                                        3-[(1,3,4-thiadiazol-5-ylthio)-                                                                  3-[(1,3,4-thiadiazol-5-                    acetic acid chloride                                                                          methyl]-7-amino-8-oxo-5-thia-1-                                                                  ylthio)methyl]-7-[[2-[4-                                   azabicyclo[4.2.0]oct-2-ene-2-car-                                                                (chloromethyl)phenoxy]-                                    boxylic acid       acetyl]amino]-8-oxo-5-                                                        thia-1-azabicyclo[4.2.0]-                                                     oct-2-ene-2-carboxylic                                                        acid                                       p-chloromethyldihydro-                                                                        3-[(3-methyl-1,2,4-thiadiazol-5-                                                                 3-[(3-methyl-1,2,4-thiadia-                cynnamic acid chloride                                                                        ylthio)methyl]-7-amino-8-oxo-5-                                                                  zol-5-ylthio)methyl]-7-[[3-                                thia-1-azabicyclo[4.2.0]oct-2-ene-                                                               [4-(chloromethyl)phenyl]-                                  2-carboxylic acid  propionyl]amino]-8-oxo-5-                                                     thia-1-azabicyclo[4.2.0]-                                                     oct-2-ene-2-carboxylic                                                        acid                                       4-(p-chloromethylphenyl)-                                                                     3-[(tetrazol-5-ylthio)methyl]-7-                                                                 3-[(tetrazol-5-ylthio)-                    butyric acid chloride                                                                         amino-8-oxo-5-thia-1-azabicyclo-                                                                 methyl]-7-[[4-[4-(chloro-                                  [4.2.0]oct-2-ene-2-carboxylic                                                                    methyl)phenyl]butyryl]-                                    acid               amino]-8-oxo-5-thia-1-aza-                                                    bicyclo[4.2.0]oct-2-ene-2-                                                    carboxylic acid                            p-chloromethylphenylacetyl                                                                    3-[(1-methyltetrazol-5-ylthio)-                                                                  3-[(1-methyltetrazol-5-yl-                 chloride        methyl]-7-amino-8-oxo-5-thia-1-                                                                  thio)methyl]-7-[[2-[4-                                     azabicyclo[4.2.0]oct-2-ene-2-                                                                    (chloromethyl)phenyl]ace-                                  carboxylic acid    tyl]amino]-8-oxo-5-thia-1-                                                    azabicyclo[4.2.0]oct-2-ene-                                                   2-carboxylic acid                          4-(p-chloromethylphenoxy)-                                                                    3-[(2-methyl-1,3,4-oxadiazol-5-                                                                  3-[(2-methyl-1,3,4-oxadia-                 butyric acid chloride                                                                         ylthio)methyl]-7-amino-8-oxo-5-                                                                  zol-5-ylthio)methyl]-7-[[4-                                thia-1-azabicyclo[4.2.0]oct-2-                                                                   [4-(chloromethyl)phenyl]-                                  ene-2-carboxylic acid                                                                            butyryl]amino]-8-oxo-5-thia-                                                  1-azabicyclo[4.2.0]oct-2-                                                     ene-2-carboxylic acid                      3-(p-chloromethylphenoxy)-                                                                    3-[(3-methyl-1,2,4-thiadiazol-                                                                   3-[(3-methyl-1,2,4-thiadia-                propionic acid  5-ylthio)methyl]-7-amino-8-oxo-                                                                  zol-5-ylthio)methyl]-7-[[3-                                5-thia-1-azabicyclo[4.2.0]oct-                                                                   [4-(chloromethyl)phenoxy]-                                 2-ene-2-carboxylic acid propionyl-                                                               propionyl]amino]-8-oxo-5-                                  oxymethyl ester    thia-1-azabicyclo[4.2.0]-                                                     oct-2-ene-2-carboxylic                                                        acid propionyloxymethyl                                                       ester                                      2-(p-chloromethyl)anilino-                                                                    3-[(1-methyltetrazol-5-ylthio)-                                                                  3-[(1-methyltetrazol-5-yl-                 propionic acid chloride                                                                       methyl]-7-amino-8-oxo-5-thia-1-                                                                  thio)methyl]-7-[[2-[4-                                     azabicyclo[4.2.0]oct-2-ene-2                                                                     (chloromethyl)anilino]-2-                                  carboxylic acid pivalyloxymethyl                                                                 methylacetyl]amino]-8-oxo-                                 ester              5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                                   pivalyloxymethyl ester                     3-(p-chloromethylanilino)-                                                                    3-[(1,3,4-thiadiazol-5-ylthio)-                                                                  3-[(1,3,4-thiadiazol-5-                    butyric acid chloride                                                                         methyl]-7-amino-8-oxo-5-thia-1-                                                                  ylthio)methyl]-7-[[3-[4-                                   azabicyclo[4.2.0]oct-2-ene-2-                                                                    (chloromethyl)anilino]-2-                                  carboxylic acid acetyloxymethyl                                                                  methylpropionyl]amino]-8-                                  ester              oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-carbox-                                                    ylic acid acetyloxymethyl                                                     ester                                      2-(p-chloromethylphenyl)-                                                                     3-[(tetrazol-5-ylthio)methyl]-7-                                                                 3-[(tetrazol-5-ylthio)methyl]-             thiopropionic acid chloride                                                                   amino-8-oxo-5-thia-1-azabicyclo-                                                                 7-[[2-[4-(chloromethyl)phen-                               [4.2.0]oct-2-ene-2-carboxylic                                                                    ylthio]-2-methylacetyl]-                                   acid butyryloxymethyl ester                                                                      amino]-8-oxo-5-thia-1-azabi-                                                  cyclo[4.2.0]oct-2-ene-2-                                                      carboxylic acid butyryloxy-                                                   methyloxymethyl ester                      o-chloro-p-chloromethyl-                                                                      3-[(2-methyl-1,3,4-oxadiazol-5-                                                                  3-[(2-methyl-1,3,4-oxadia-                 phenylacetic acid chloride                                                                    ylthio)methyl]-7-amino-8-oxo-5-                                                                  zol-5-ylthio)methyl]-7-[[2-                                thia-1-azabicyclo[4.2.0]oct-2-                                                                   [2-(chloro)-4-(chloromethyl)-                              ene-2-carboxylic acid 2-amino-3-                                                                 phenyl]acetyl]amino]-8-oxo-                                methylbutyryloxymethyl ester                                                                     5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                                                   2-amino-3-methylbutyryloxy-                                                   methyl ester                               2-chloromethyl-4-methoxy-                                                                     2-[(2-methyl-1,3,4-thiadiazol-                                                                   3-[(2-methyl-1,3,4-thia-                   mandelic acid chloride                                                                        5-ylthio)methyl]-7-amino-8-oxo-                                                                  diazol-5-ylthio)methyl]-                                   5-thia-1-azabicyclo[4.2.0]oct-                                                                   7-[[2-[2-(chloromethyl)-                                   2-ene-2-carboxylic acid N-ethoxy-                                                                4-methoxypnenyl]-2-hydroxy-                                carbonyl-N-methylaminomethyl                                                                     acetyl]amino]-8-oxo-5-thia-                                ester              1-azabicyclo[4.2.0]oct-2-                                                     ene-2-carboxylic acid N-                                                      ethoxycarbonyl-N-methyla-                                                     minomethyl ester                           3-(p-chloromethylphenyl)-                                                                     3-[(1-methyltetrazol-5-ylthio)-                                                                  3-[(1-methyltetrazol-5-yl-                 lactic acid chloride                                                                          methyl]-7-amino-8-oxo-5-thia-1-                                                                  thio)methyl]-7-[[3-[4-                                     azabicyclo[4.2.0]oct-2-ene-2-                                                                    (chlorometyl)phenyl]-2-                                    carboxylic acid p-pivalyloxy-                                                                    hydroxypropionyl]amino]-8-                                 benzyl ester       oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-carbox-                                                    ylic acid p-pivalyloxy-                                                       benzyl ester                               __________________________________________________________________________

EXAMPLE 197-Amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester

To 35 ml of dimethylformamide is added 7.5 g of the sodium salt of7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and the solution is stirred at room temperature for about 30minutes after which 8 ml of chloromethylpivalate is added. Stirring iscontinued at room temperature for about 3 hours. The mixture is dilutedwith ethylacetate and washed with water. The organic layer is separated,evaporated to dryness, and the residue is recrystallized fromethylacetateto give7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester.

In a similar manner when an appropriate amount ofchloromethylpropionate, chloromethylacetate or chloromethylbutyrate issubstituted for chloromethylpivalate in the above procedure, thefollowing respective products are obtained.

7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester

7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid acetyloxymethyl ester

7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid butyryloxymethyl ester.

When in the procedure of Example 16 an appropriate amount of7-amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid or the corresponding pivalyloxymethyl ester is substituted for3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid,3-methyl-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and3-methyl-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester are obtained.

EXAMPLE 203-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

p-Chloromethylphenylglycine wherein the amino group is protected withtert-butoxycarbonyl is treated with isobutylchloroformate in thepresence of triethylamine. The thus obtained anhydride is reacted withthe triethylamine salt of3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid at 0° C. for about 4 hours.The resulting product is isolated, andthe amine protecting group is removed by acid hydrolysis to give3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-[[2-[4-chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

When appropriate amounts of an amino acid derivative and a7-aminocephalosporanic acid derivative listed in the following Table IXare substituted respectively for p-chloromethylphenylglycine and3-[(2-methyl-1,3,4-thiadiazol-5-yl-thio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in the above procedure, the respective products listed in thefollowing Table IX are obtained.

                                      TABLE IX                                    __________________________________________________________________________                  7-AMINOCEPHALOSPORANIC                                          AMINO ACID    ACID DERIVATIVE   PRODUCT                                       __________________________________________________________________________    3-(p-chloromethylphenyl)-                                                                   3-[(1,3,4-thiadiazol-5-ylthio)-                                                                 3-[(1,3,4-thiadiazol-                         alanine       methyl]-7-amino-8-oxo-5-thia-1-                                                                 5-ylthio)methyl]-7-[[3-                                     azabicyclo[4.2.0]oct-2-ene-2-                                                                   [4-(chloromethyl)phenyl]-                                   carboxylic acid   2-aminopropionyl]amino]-                                                      8-oxo-5-thia-1-azabicy-                                                       cyclo[4.2.0]oct-2-ene-2-                                                      carboxylic acid                               2-amino-4-(p-chloromethyl-                                                                  3-[(3-methyl-1,2,4-oxadiazol-                                                                   3-[(3-methyl-1,2,4-thia-                      phenoxy)butyric acid                                                                        5-ylthio)methyl]-7-amino-8-                                                                     diazol-5-ylthio)methyl]-7-                                  oxo-5-thia-1-azabicyclo[4.2.0]-                                                                 [[4-[4-(chloromethylphen-                                   oct-2-ene-2-carboxylic acid                                                                     oxy]-2-aminobutyryl]amino]-                                                   8-oxo-5-thia-1-azabicy-                                                       clo[4.2.0]oct-2-ene-2-car-                                                    boxylic acid                                  3-(p-chloromethylphenoxy)-                                                                  3-[(tetrazol-5-ylthio)methyl]-                                                                  3-[(tetrazol-5-ylthio)-                       alanine       7-amino-8-oxo-5-thia-1-azabi-                                                                   methyl]-7-[[3-[4-(chloro-                                   cyclo[4.2.0]oct-2-ene-2-car-                                                                    methyl]phenoxy]-2-amino-                                    boxylic acid      propionyl]amino]-8-oxo-5-                                                     thia-1-azabicyclo[4.2.0]-                                                     oct-2-ene-2-carboxylic                                                        acid                                          2-amino-4-(p-chloromethyl-                                                                  3-[(1-methyltetrazol-5-ylthio)-                                                                 3-[(1-methyltetrazol-5-                       phenyl)thiobutyric acid                                                                     methyl]-7-amino-8-oxo-5-thia-1-                                                                 ylthio)methyl]-7-[[4-[4-                                    azabicyclo[4.2.0]oct-2-ene-2-                                                                   (chloromethyl)phenylthio]-                                  carboxylic acid   2-aminobutyryl]amino]-8-                                                      oxo-5-thia-1-azabicyclo-                                                      [4.2.0]oct-2-ene-2-car-                                                       boxylic acid                                  2-[2-(5-chloromethyl)-                                                                      3-[(2-methyl-1,3,4-oxadiazol-5-                                                                 3-[(2-methyl-1,3,4-oxa-                       thienyl]glycine                                                                             ylthio)methyl]-7-amino-8-oxo-5-                                                                 diazol-5-ylthio)methyl]-                                    thia-1-azabicyclo[4.2.0]oct-2-                                                                  7-[[2-[5-(chloromethyl)-2-                                  ene-2-carboxylic acid                                                                           thienyl]-2-aminoacetyl]-                                                      amino]-8-oxo-5-thia-1-                                                        azabicyclo[4.2.0]oct-2-                                                       ene-2-carboxylic acid                         __________________________________________________________________________

EXAMPLE 213-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethylester

p-Chloromethylphenylglycine wherein the amino group is protected withtert-butoxycarbonyl is treated with isobutyl chloroformate in thepresenceof triethylamine. The thus obtained mixed anhydride is reactedwith3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester at 0° C. for about 4 hours. The resultingproduct is isolated, and the amine protecting group is removed by acidhydrolysis by treating the product with difluoroacetic acid for 5minutes at room temperature to give3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester.

When in the above procedure an appropriate amount of3-[(1-methyltetrazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester is substituted for3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester, the following compound is obtained:

7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-3-[(1-methyltetrazol-5-ylthio)methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid pivalyloxymethyl ester

EXAMPLE 227-[[2-[4-(Chloromethyl)phenyl]-2-carboxyacetyl]amino]-3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

α-Carboxy-p-chloromethylphenylacetylnitrophenyl polymer, preparedaccording to the procedure described in Canadian Pat. No. 892,580,carrying 4m. mole of p-chloromethylphenylmalonic acid is suspended forabout 8 hours in 20 ml of dry methylene chloride solution containing 1m. mole of3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid triethylammonium salt, which is prepared from the7-aminocephalosporanic acid derivative and triethylamineat roomtemperature. After only traces of3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid remain in solution, which is determined by thin layerchromatography on cellulose in 70% aqueous propanol, the polymer isfiltered off and washed with three portions of 50ml each of methylenechloride. The combined filtrates are evaporated and the residue isdissolved in 20 ml of distilled water. This solution is acidified to pH2 by adding 0.2 N hydrochloric acid and extracted twice withethylacetate. The organic solution is dried over sodium sulfate andevaporated to room temperature. The remaining solid is dried overnightover phosphorus pentoxide under vacuum to give7-[[2-[4-chloromethyl)phenyl]-2-carboxyacetyl]amino]-3-[(2-methyl-1,3,4-thiadiazol-5-yl-thio)methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

When in the above procedure appropriate amounts of an acid derivativeand acephalosporin derivative listed in the following Table X aresubstituted respectively for p-chloromethylphenylmalonic acid and3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in the above procedure, the respective products listed in thefollowing Table X are obtained.

                                      TABLE X                                     __________________________________________________________________________    ACID DERIVATIVE                                                                             CEPHALOSPORIN DERIVATIVE                                                                         PRODUCT                                      __________________________________________________________________________    2-sulfo-p-chloromethyl-                                                                     3-[(1,3,4-thiadiazol-5-ylthio)-                                                                  7-[[2-[4-(chloromethyl)-                     phenylacetic acid                                                                           methyl]-7-amino-8-oxo-5-thia-1-                                                                  phenyl]-2-sufloacetyl[-                                    azabicyclo[4.2.0]oct-2-ene-2-car-                                                                amino]-3-[(1,3,4-thiadia-                                  boxylic acid       zol-5-ylthio)methyl]-8-oxo-                                                   5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic acid                  4-(p-chloromethylphenyl)-                                                                   3-[(3-methyl-1,2,4-thiadiazol-                                                                   7-[[4-[4-(chloromethyl)phenyl]-              2-sulfobutyric acid                                                                         5-ylthio)methyl]-7-amino-8-oxo-                                                                  2-sulfobutyryl]amino]-3-[(3-                               5-thia-1-azabicyclo[4.2.0]oct-                                                                   methyl-1,2,4-thiadiazol-5-yl-                              2-ene-2-carboxylic acid                                                                          thio)methyl]-8-oxo-5-thia-1-                                                  azabicyclo[4.2.0]oct-2-ene-                                                   2-carboxylic acid                            2-(p-chloromethylphen-                                                                      3-[(tetrazol-5-ylthio)methyl]-                                                                   7-[[4-[4-(chloromethyl)phen-                 oxy)ethylmalonic acid                                                                       7-amino-8-oxo-5-thia-1-azabi-                                                                    oxy]-2-carboxybutyryl]amino]-                              cyclo[4.2.0]oct-2-ene-2-car-                                                                     3-[(tetrazol-5-ylthio)methyl]-                             boxylic acid       8-oxo-5-thia-1-azabicyclo-                                                    [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                         p-chloromethylanilino-                                                                      3-[(1-methyltetrazol-5-ylthio)-                                                                  7-[[3-[4-(chloromethyl)ani-                  methylmalonic acid                                                                          methyl]-7-amino-8-oxo-5-thia-1-                                                                  lino]-2-carboxypropionyl]-                                 azabicyclo[4.2.0]oct-2-ene-2-                                                                    amino]-3-[(1-methyltetrazol-                               carboxylic acid    5-ylthio)methyl]-8-oxo-5-                                                     thia-1-azabicyclo[4.2.0]oct-                                                  2-ene-2-carboxylic acid                      2-[2-(5-chloromethyl)-                                                                      3-[(2-methyl-1,3,4-oxadiazol-                                                                    7-[[3-[5-(chloromethyl)-2-                   thenyl]malonic acid                                                                         5-ylthio)methyl]-7-amino-8-                                                                      thienyl]-2-carboxypropionyl]-                              oxo-5-thia-1-azabicyclo[4.2.0]-                                                                  amino]-3-[(2-methyl-1,3,4-oxa-                             oct-2-ene-2-carboxylic acid                                                                      diazol-5-ylthio)methyl]-8-oxo-                                                5-thia-1-azabicyclo[4.2.0]-                                                   oct-2-ene-2-carboxylic                       __________________________________________________________________________                                     acid                                     

EXAMPLE 233-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester

α-Carboxy-p-chloromethylphenylacetylnitrophenyl polymer prepared as inthe procedure of Example 22, carrying four m. mole ofp-chloromethylphenylmalonic acid was suspended for about 8 hours in 20ml of dry methylene chloride solution containing 1 millimole of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester. After only traces of the7-aminocephalosporanic acid ester derivative remain in solution, whichis determined by thin layer chromatography on cellulose in 70% aqueouspropanol, the polymer is filtered off and washed with portions of 50 mleach of methylene chloride. The combined filtrates are evaporated andthe residue is dissolved in 20 ml of distilled water. The solution isacidified to a pH of 2 by adding 0.2 normal hydrochloric acid andextracted twice with ethylacetate. The organic solution is dried oversodium sulfate and evaporated at room temperature. The remaining solidis dried overnight over phosphorus pentoxide under vacuum to give3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acidpropionyloxymethyl ester.

When in the above procedure an appropriate amount of7-amino-3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester is substituted for3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid propionyloxymethyl ester the following product is obtained:

7-[[2-[4-(chloromethyl)phenyl]-2-carboxyacetyl]amino]-3-[(2-methyl-1,3,4-thiadiazol-5-ylthio)methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

EXAMPLE 243-[(Acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-7-methoxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

When in the procedure of Example 14 an appropriate amount of3-[(acetyloxy)methyl]-7-amino-7-methoxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid is substituted for3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid the title compound is obtained.

In a similar manner when an appropriate amount of3-[(acetyloxy)methyl]-7-amino-7-methoxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid is reacted with the acid chloride listed in TableV the 7-methoxyderivatives of the respective cephalosporin derivatives listed in TableV are obtained.

EXAMPLE 253-[(Acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

A mixture of 0.3 g of3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in 10 ml of pyridine was stirred at room temperature for 24 hoursafter which the excess pyridine was removed under high vacuum at roomtemperature yielding a yellow powder which was triturated with absoluteethanol to give3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid. M.P. 102° C. (dec.).

EXAMPLE 263-Methyl-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

When in the procedure of Example 25 0.3 g of7-[[2-[4-(chloromethyl)phenyl]acetyl]amino-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicis substituted for3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid the following product is obtained:

3-methyl-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid M.P. 102° C. (dec.).

When in the procedure of Example 25 an appropriate amount of achloromethylsubstituted cephalosporin derivative listed in the followingTable XI is substituted for3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid the respective pyridiniummethyl substituted cephalosporin productlisted in Table XI is obtained.

                  TABLE XI                                                        ______________________________________                                        CHLOROMETHYL SUBSTI-                                                                           PYRIDINIUMMETHYL                                             TUTED CEPHALOSPORIN                                                                            SUBSTITUTED                                                  DERIVATIVE       CEPHALOSPORIN PRODUCT                                        ______________________________________                                        3-[(acetyloxy)methyl-7-                                                                        3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-methylacetyl]-                                                                       2-methylacetyl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-a-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]propionyl]amino]-                                                                       propionyl]amino]-8-oxo-5-                                    8-oxo-5-thia-1-azabicy-                                                                        thia-1-azabicyclo[4.2.0]oct-                                 clo[4.2.0]oct-2-ene-2-                                                                         2-ene-2-carboxylic acid                                      carboxylic acid                                                               3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [-(pyridiniummethyl)phenyl]-                                 phenyl]-2-methylpropionyl]-                                                                    2-methylpropionyl]amino]-8-                                  amino]-8-oxo-5-thia-1-                                                                         oxo-5-thia-1-azabicyclo[4.2.0]-                              azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl]phenyl]-                                phenyl]butyryl]amino]-8-                                                                       butyryl]amino]-8-oxo-5-thia-                                 oxo-5-thia-1-azabicyclo-                                                                       1-azabicyclo[4.2.0]oct-2-ene-                                [4.2.0]oct-2-ene-2-car-                                                                        2-carboxylic acid                                            boxylic acid                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-methylbutyryl]-                                                                      2-methylbutyryl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]  acetyl]amino]-8-                                                                     acetyl]amino]-8-oxo-5-thia-                                  oxo-5-thia-1-azabicyclo-                                                                       1-azabicyclo[4.2.0]oct-2-ene-                                [4.2.0]oct-2-ene-2-car-                                                                        2-carboxylic acid                                            boxylic acid                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-methylacetyl]-                                                                      2-methylacetyl]amino]-8-oxo-                                 amino]8-oxo-5-thia-1-                                                                          5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl]-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]butyryl]amino]-8-                                                                       butyryl]amino]-8-oxo-5-thia-                                 oxo-5-thia-1-azabicyclo-                                                                       1-azabicyclo[4.2.0]oct-2-ene-                                [4.2.0]oct-2-ene-2-car-                                                                        2-carboxylic acid                                            boxylic acid                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-methylbutyryl]-                                                                     2-methylbutyryl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]propionyl]amino]-                                                                      propionyl]amino]-8-oxo-5-                                    8-oxo-5-thia-1-azabicy-                                                                        thia-1-azabicyclo[4.2.0]oct-                                 clo[4.2.0]oct-2-ene-2-                                                                         2-ene-2-carboxylic acid                                      carboxylic acid                                                               3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]acetyl]amino]-                                                                         acetyl]amino]-8-oxo-5-thia-                                  8-oxo-5-thia-1-azabicy-                                                                        1-azabicyclo[4.2.0]oct-2-ene-                                clo[4.2.0]oct-2-ene-2-                                                                         2-carboxylic acid                                            carboxylic acid                                                               3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]  -2-methylacetyl]-                                                                    2-methylacetyl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]butyryl]amino]-                                                                        butyryl]amino]-8-oxo-5-thia-                                 8-oxo-5-thia-1-azabi-                                                                          1-azabicyclo[4.2.0]oct-2-ene-                                cyclo[4.2.0]oct-2-ene-                                                                         2-carboxylic acid                                            2-carboxylic acid                                                             3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]-2-methylpro-                                                                          2-methylpropionyl]amino]-8-                                  pionyl]amino]-8-oxo-5-                                                                         oxo-5-thia-1-azabicyclo[4.2.0]-                              thia-1-azabicyclo[4.2.0]-                                                                      oct-2-ene-2-carboxylic acid                                  oct-2-ene-2-carboxylic                                                        acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)]-                                                                        [4-(pyridiniummethyl)phenyl-                                 phenylthio]acetyl]amino]-                                                                      thio]acetyl]amino]-8-oxo-5-                                  8-oxo-5-thia-1-azabicy-                                                                        thia-1-azabicyclo[4.2.0]oct-                                 clo[4.2.0]oct-2-ene-2-                                                                         2-ene-2-carboxylic acid                                      carboxylic acid                                                               3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]7-[[2-                                  [[2-[4-(chloromethyl)- [4-(pyridiniummethyl)phenyl-                           phenylthio]-2-methyl-                                                                          thio]-2-methylacetyl]amino]-                                 acetyl]amino]-8-oxo-5-                                                                         8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]butyryl]-                                                                           thio]butyryl]amino]-8-oxo-5-                                 amino]-8-oxo-5-thia-1-                                                                         thia-1-azabicyclo[4.2.0]oct-                                 azabicyclo[4.2.0]oct-                                                                          2-ene-2-carboxylic acid                                      2-ene-2-carboxylic acid                                                       3-[(acetyloxy)methyl[-7-                                                                       3-[(acetyloxy)methyl]-7-[[ 2-                                [[2-[2-(chloro)-4-(chloro-                                                                     [2-(chloro)-4-(pyridinium- -methyl]phenyl]acetyl]- methyl                     )phenyl]acetyl]amino]-                                       amino]-8-oxo-5-thia-1-                                                                         8-oxo-5-thia-1-azabicyclo-                                   azabicyclo[4.2.0]oct-2-                                                                        [4.2.0]oct-2-ene-2-carboxylic                                ene-2-carboxylic acid                                                                          acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-8-[[2-                                 [[2-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thi-                                 thienyl]acetyl]amino]-8-                                                                       enyl]-acetyl]amino]-8-oxo-5-                                 oxo-5-thia-1-azabicyclo-                                                                       thia-1-azabicyclo[4.2.0] oct-                                [4.2.0]oct-2-ene-2-car-                                                                        2-ene-2-carboxylic acid                                      boxylic acid                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[2-(chloromethyl)-4-                                                                       [2-pyridiniummethyl-4-                                       methoxyphenyl]-2-hydroxy-                                                                      methoxyphenyl]-2-hydroxyace-                                 acetyl]amino]-8-oxo-5-                                                                         tyl]amino]-8-oxo-5-thia-1-                                   thia-1-azabicyclo[4.2.0]-                                                                      azabicyclo[4.2.0]oct-2-ene-2-                                oct-2-ene-2-carboxylic                                                                         carboxylic acid                                              acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl] -                               phenyl]-2-hydroxyacetyl]-                                                                      2-hydroxyacetyl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7[[3-                                  [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-hydroxypropi-                                                                        2-hydroxypropionyl]amino]-8-                                 onyl]amino]-8-oxo-5-thia-                                                                      oxo-5-thia-1-azabicyclo-                                     1-azabicyclo[4.2.0]oct-                                                                        [4.2.0]oct-2-ene-carbox-                                     2-ene-2-carboxylic acid                                                                        ylic acid                                                    3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-hydroxybutyryl]-                                                                     2-hydroxybrutyryl]amino]-8-                                  amino]-8-oxo-5-thia-1-                                                                         oxo-5-thia-1-azabicyclo-                                     azabicyclo[4.2.0]oct-2-                                                                        [4.2.0]oct-2-ene-2-carbox-                                   ene-2-carboxylic acid                                                                          ylic acid                                                    3-[(acetyloxy)methyl[-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-hydroxypro-                                                                         2-hydroxypropionyl]amino]-8-                                 pionyl]amino]-8-oxo-5-                                                                         oxo-5-thia-1-azabicyclo-                                     thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]- 7-[[2-                                [[2-[5-(chloromethyl)-                                                                         [5-(pyridiniummethyl)-2-thi-                                 2-thienyl]-2-hydroxy-                                                                          enyl]-2-hydroxyacetyl]amino]                                 acetyl]amino]-8-oxo-5-                                                                         8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0] oct-2-ene-2-carboxylic                               oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[(acetyloxy)methyl[-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-carboxyacetyl]-                                                                      2-carboxyacetyl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-sulfoacetyl]-                                                                        2-sulfoacetyl]amino]-8-oxo-                                  amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]oct-                               azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl[-7-[ [4-                                [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-sulfobutyryl]  -                                                                     2-sulfobutyryl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]oct-                               azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-carboxypropi-                                                                        2-carboxypropionyl]amino]-8-                                 onyl]amino]-8-oxo-5-thia-                                                                      oxo-5-thia-1-azabicyclo[4.2.0]-                              1-azabicyclo[4.2.0]oct-                                                                        oct-2-ene-2-carboxylic acid                                  2-ene-2-carboxylic acid                                                       3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl)-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-carboxybutyryl]-                                                                    2-carboxybutyryl]amino]-8-oxo-                               amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]oct-                               azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]-2-carboxy-                                                                         thio]-2-carboxybutyryl]amino]-                               butyryl]amino]-8-oxo-5-                                                                        8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                        acid             acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]- 7[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]-2-carboxypro-                                                                         2-carboxypropionyl]amino]-8-                                 pionyl]amino]-8-oxo-5-                                                                         oxo-5-thia-1-azabicyclo[4.2.0]-                              thia-1-azabicyclo-                                                                             oct-2-ene-2-carboxylic acid                                  [4.2.0]oct-2-ene-2-                                                           carboxylic acid                                                               3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thi-                                 thienyl]-2-carboxypro-                                                                         enyl]-2-carboxypropionyl]-                                   pionyl]amino]-8-oxo-5-                                                                         amino]-8-oxo-5-thia-1-azabi-                                 thia-1-azabicyclo[4.2.0]-                                                                      cyclo[4.2.0]oct-2-ene-2-car-                                 oct-2-ene-2-carboxylic                                                                         boxylic acid                                                 acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]  -7-[[2-                               [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]acetyl]amino]-8-                                                                        acetyl]amino]-8-oxo-5-thia-1-                                oxo-5-thia-1-azabicyclo-                                                                       azabicyclo[4.2.0]oct-2-ene-2-                                [4.2.0]oct-2-ene-2-car-                                                                        carboxylic acid pivalyloxy-                                  boxylic acid     methyl ester                                                 3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-methylacetyl]-                                                                       2-methylacetyl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                                          propionyloxymethyl ester                                     propionyloxymethyl ester                                                      3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-methylpropionyl]-                                                                    2-methylpropionyl]amino]-8-                                  amino]-8-oxo-5-thia-1-                                                                         oxo-5-thia-1-azabicyclo-                                     azabicyclo[4.2.0]oct-2-                                                                        [4.2.0]oct-2-ene-2-carboxylic                                ene-2-carboxylic acid ace-                                                                     acid acetyloxymethyl ester                                   tyloxymethyl ester                                                            3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-methylbutyryl]-                                                                      2-methylbutyryl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                                          butyryloxymethyl ester                                       butyryloxymethyl ester                                                        3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenoxy]-2-methylacetyl]-                                                                      2-methylacetyl]amino]-8-oxo                                  amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid 2-                                                                       2-amino-3-methylbutyryloxy-                                  amino-3-methylbutyryloxy-                                                                      methyl ester                                                 methyl ester                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-pyridiniummethyl)phenyl]-                                 phenoxy]-2-methylbutyryl]-                                                                     2-methylbutyryl]amino]-8-oxo                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid N-                                                                       N-ethoxycarbonyl-N-methyl-                                   ethoxycarbonyl-N-methyl-                                                                       aminoethyl ester                                             aminoethyl ester                                                              3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]acetyl]amino]-8-                                                                       acetyl]amino]-8-oxo-5-thia-1-                                oxo-5-thia-1-azabicyclo-                                                                       azabicyclo[4.2.0]oct-2-ene-                                  [4.2.0]oct-2-ene-2-car-                                                                        2-carboxylic acid N-methyl-                                  boxylic acid N-methyl-N-                                                                       N-propionylaminoethyl ester                                  propionylaminomethyl                                                          ester                                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]butyryl]amino]-                                                                        butyryl]amino]-8-oxo-5-thia-                                 8-oxo-5-thia-1-azabi-                                                                          1-azabicyclo[4.2.0]oct-2-ene-                                cyclo[4.2.0]oct-2-ene-2-                                                                       2-carboxylic acid N-butyryl-                                 carboxylic acid N-butyryl-                                                                     aminomethyl ester                                            aminomethyl ester                                                             3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]acetyl]amino]-                                                                      thio]acetyl]amino]-8-oxo-5-                                  8-oxo-5-thia-1-azabi-                                                                          thia-1-azabicyclo[4.2.0]oct-                                 cyclo[4.2.0]oct-2-ene-2-                                                                       2-ene-2-carboxylic acid N-                                   carboxylic acid N-acetyl-                                                                      acetylaminomethyl ester                                      aminomethyl ester                                                             3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]butyryl]amino]-                                                                     thio]butyryl]amino]-8-oxo-                                   8-oxo-5-thia-1-azabicy-                                                                        5-thia-1-azabicyclo[4.2.0]-                                  clo[4.2.0]oct-2-ene-2-car-                                                                     oct-2-ene-2-carboxylic acid                                  boxylic acid N-methyl-N-                                                                       N-methyl-N-ethoxycarbonyl-                                   ethoxycarbonylaminomethyl                                                                      aminomethyl ester                                            ester                                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thi-                                 thienyl]acetyl]amino]-8-                                                                       enyl]acetyl]amino]-8-oxo-5-                                  oxo-5-thia-1-azabicyclo-                                                                       thia-1-azabicyclo[4.2.0]oct-                                 [4.2.0]oct-2-ene-2-car-                                                                        2-ene-2-carboxylic acid p-                                   boxylic acid p-pivalyloxy-                                                                     pivalyloxybenzyl ester                                       benzyl ester                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-hydroxyacetyl]-                                                                      2-hydroxyacetyl]amino]-8-oxo-                                amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                                          p-(propionyloxy)benzyl ester                                 p-(propionyloxy)benzyl                                                        ester                                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-hydroxybutyryl]-                                                                     2-hydroxybutyryl]amino]-8-oxo-                               amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid p-                                                                       p-(acetyloxy)benzyl ester                                    (acetyloxy)benzyl ester                                                       3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thi-                                 thienyl]-2-hydroxyacetyl]-                                                                     enyl]-2-hydroxyacetyl]amino]-                                amino]-8-oxo-5-thia-1-aza-                                                                     8-oxo-5-thia-1-azabicyclo-                                   bicyclo[4.2.0]oct-2-ene-                                                                       [4.2.0]oct-2-ene-2-carboxylic                                2-carboxylic acid p-(val-                                                                      acid p-valyryloxy)benzyl ester                               yryloxy)benzyl ester                                                          3-[(2-methyl-1,3,4-thia-                                                                       3-[(2-methyl-1,3,4-thiadiazol-                               diazol-5-ylthio)methyl]-7-                                                                     5-ylthio)methyl]-7-[ [2-[4-                                  [[2-[4-(chloromethyl)phen-                                                                     pyridiniummethyl)phenyl]acetyl]                              yl]acetyl]amino]-8-oxo-5-                                                                      amino]-8-oxo-5-thia-1-azabi-                                 thia-1-azabicyclo[4.2.0]-                                                                      cyclo[4.2.0]oct-2-ene-2-car-                                 oct-2-ene-2-carboxylic                                                                         boxylic acid                                                 acid                                                                          3-[(1,3,4-thiadiazol-5-                                                                        3-[(1,3,4-thiadiazol-5-ylthio)-                              ylthio)methyl]-7-[[2[4-                                                                        methyl]-7-[[2-[4-pyridinium-                                 (chloromethyl)phenoxy]-                                                                        methyl)phenoxypacetyl]amino]-                                acetyl]amino]-8-oxo-5-                                                                         8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[(3-methyl-1,2,4-thia-                                                                       3-[(3-methyl-1,2,4-thiadiazol-                               diazol-5-ylthio)methyl]-                                                                       5-ylthio)methyl]-7-[[3-[4-                                   7-[[3-[4-(chloromethyl)-                                                                       (pyridiniummethyl)phenyl]pro-                                phenyl]propionyl]amino]-                                                                       pionyl]amino]-8-oxo-5-thia-1-                                8-oxo-5-thia-1-azabi-                                                                          azabicyclo[4.2.0]oct-2-ene-2-                                cyclo[4.2.0]oct-2-ene-2-                                                                       carboxylic acid                                              carboxylic acid                                                               3-[(tetrazol-5-ylthio)-                                                                        3-[(tetrazol-5-ylthio)methyl]-                               methyl]-7-[[4-[4-(chloro-                                                                      7-[[4-[4-(pyridiniummethyl)-                                 methyl)phenyl]butyryl]-                                                                        phenyl]butyryl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-aza-                                                                     5-thia-1-azabicyclo[4.2.0]-                                  bicyclo[4.2.0]oct-2-ene-2-                                                                     oct-2-ene-2-carboxylic acid                                  carboxylic acid                                                               3-[(1-methyltetrazol-5-yl-                                                                     3-[(1-methyltetrazol-5-ylthio)-                              thio)methyl]-7-[[2-[4-                                                                         methyl]-7-[[2-[4-(pyridinium-                                (chloromethyl)phenyl]ace-                                                                      methyl)phenyl]acetyl]amino]-                                 tyl]amino]-8-oxo-5-thia-1-                                                                     8-oxo-5-thia-1-azabicyclo-                                   azabicyclo[4.2.0]oct-2-ene-                                                                    [4.2.0]oct-2-ene-2-carboxylic                                2-carboxylic acid                                                                              acid                                                         3-[(2-methyl-1,3,4-oxadia-                                                                     3-[(2-methyl-1,3,4-oxadiazol-                                zol-5-ylthio)methyl]-7-[[4-                                                                    5-ylthio)methyl[-7-[[4-[4-                                   [4-(chloromethyl)phenyl]-                                                                      (pyridiniummethyl)phenyl]buty-                               butyryl]amino]-8-oxo-5-                                                                        ryl]amino]-8-oxo-5-thia-1-aza-                               thia-1-azabicyclo[4.2.0]-                                                                      bicyclo[4.2.0]oct-2-ene-2-car-                               oct-2-ene-2-carboxylic                                                                         boxylic acid                                                 acid                                                                          3-[3-methyl-1,2,4-thia-                                                                        3-[(3-methyl-1,2,4-thiadia-                                  diazol-5-ylthio)methyl]-                                                                       zol-5-ylthio)methyl]-7-[[3-                                  7-[[3-[4-(chloromethyl)-                                                                       [4-(pyridiniummethyl)phenoxy]  -                             phenoxy]propionyl]amino]-                                                                      propionyl]amino]-8-oxo-5-thia-                               8-oxo-5-thia-1-azabicy-                                                                        1-azabicyclo[4.2.0]oct-2-ene-                                clo[4.2.0]oct-2-ene-2-                                                                         2-carboxylic acid propionyl-                                 carboxylic acid propion-                                                                       oxymethyl ester                                              yloxymethyl ester                                                             3-[(1-methyltetrazol-5-                                                                        3-[(1-methyltetrazol-5-yl-                                   ylthio)methyl]-7-[[2-[4-                                                                       thio)methyl]-7-[[2-[4-(pyri-                                 (chloromethyl)anilino]-2-                                                                      diniummethyl)anilino]-2-                                     methylacetyl]amino]-8-                                                                         methylacetyl]amino]-8-oxo-                                   oxo-5-thia-1-azabicyclo-                                                                       5-thia-1-azabicyclo[4.2.0]-                                  [4.2.0]oct-2-ene-2-car-                                                                        oct-2-ene-2-carboxylic acid                                  boxylic acid pivalyloxy-                                                                       pivalyloxymethyl ester                                       methyl ester                                                                  3-[(1,3,4-thiadiazol-5-                                                                        3-[(1,3,4-thiadiazol-5-yl-                                   ylthio)methyl]-7-[[3-                                                                          thio)methyl]-7-[[3-[ 4-pyri-                                 [4-(chloromethyl)ani-                                                                          diniummethyl)anilino]-2-                                     lino]-2-methylpropionyl]-                                                                      methylpropionyl]amino]-8-                                    amino]-8-oxo-5-thia-1-                                                                         oxo-5-thia-1-azabicyclo-                                     azabicyclo[4.2.0]oct-2-                                                                        [4.2.0]oct-2-ene-2-carboxylic                                ene-2-carboxylic acid                                                                          acid acetyloxymethyl ester                                   acetyloxymethyl ester                                                         3-[(tetrazol-5-ylthio)-                                                                        3-[(tetrazol-5-ylthio)methyl]-                               methyl]-7-[[2-[4-(chloro-                                                                      7-[[2-[4-(pyridiniummethyl)-                                 methyl)phenylthio]-2-                                                                          phenylthio]-2-methylacetyl]-                                 methylacetyl]amino]-8-                                                                         amino]-8-oxo-5-thia-1-azabi-                                 oxo-5-thia-1-azabicy-                                                                          cyclo[4.2.0]oct-2-ene-2-car-                                 clo[4.2.0]oct-2-ene-2-                                                                         boxylic acid butyryloxy-                                     carboxylic acid butyryl-                                                                       methyl ester                                                 oxymethyloxymethyl ester                                                      3-[(2-methyl-1,3,4-oxadi-                                                                      3-[(2-methyl-1,3,4-oxadiazol-                                azol-5-ylthio)methyl]-7-                                                                       5-ylthio)methyl] -7-[[2-[ 2-                                 [[2-[2-(chloro)-4-(chloro-                                                                     (chloro)-4-(pyridiniummethyl)-                               methyl)phenyl]acetyl]amino]-                                                                   phenyl]acetyl]amino]-8-oxo-                                  8-oxo-5-thia-1-azabicyclo-                                                                     5-thia-1-azabicyclo[4.2.0]-                                  [4.2.0]oct-2-ene-2-carbox-                                                                     oct-2-ene-2-carboxylic acid                                  ylic acid 2-amino-3-methyl-                                                                    2-amino-3-methylbutyryloxy-                                  butyryloxymethyl ester                                                                         methyl ester                                                 3-[(2-methyl-1,3,4-thia-                                                                       3-[(2-methyl-1,3,4-thiadia-                                  diazol-5-ylthio)methyl]-                                                                       zol-5-ylthio)methyl]-7-[[2-                                  7-[[2-[2-(chloromethyl)-                                                                       [2-(pyridiniummethyl)-4-                                     4-methoxyphenyl]-2-hy-                                                                         methoxyphenyl]-2-hydroxy-                                    droxyacetyl]amino]-8-                                                                          acetyl]amino]-8-oxo-5-thia-                                  oxo-5-thia-1-azabicyclo-                                                                       1-azabicyclo[4.2.0]oct-2-                                    [4.2.0]oct-2-ene-2-car-                                                                        ene-2-carboxylic acid N-                                     boxylic acid N-ethoxy-                                                                         ethoxycarbonyl-N-methyla-                                    carbonyl-N-methylamino-                                                                        minomethyl ester                                             methyl ester                                                                  3-[(1-methyltetrazol-5-                                                                        3-[(1-methyltetrazol-5-yl-                                   ylthio)methyl]-7-[[3-[4-                                                                       thio)methyl]-7-[[3-[4-                                       (chloromethyl)phenyl]-2-                                                                       pyridiniummethyl)phenyl]-2-                                  hydroxypropionyl]amino]-                                                                       hydroxypropionyl]amino]-8-                                   8-oxo-5-thia-1-azabicy-                                                                        oxo-5-thia-1-azabicyclo-                                     clo[4.2.0]oct-2-ene-2-                                                                         [4.2.0]oct-2-ene-2-carboxylic                                carboxylic acid p-piva-                                                                        acid p-pivalyloxybenzyl ester                                lyloxybenzyl ester                                                            7-[[2-[4-(chloromethyl)-                                                                       7-[[2-[4-(pyridiniummethyl)-                                 phenyl]-2-(carboxyacetyl]-                                                                     phenyl]-2-carboxyacetyl]amino]-                              amino]-3-[(2-methyl-1,3,4-                                                                     3-[(methyl-1,3,4-thiadiazol-                                 thiadiazol-5-ylthio)-                                                                          5-ylthio)methyl]-8-oxo-5-thia-                               methyl]-8-oxo-5-thia-1-                                                                        1-aziabicyclo[4.2.0]oct-2-ene-                               azabicyclo[4.2.0]oct-2-                                                                        2-carboxylic acid                                            ene-2-carboxylic acid                                                         7-[[2-[4-(chloromethyl)-                                                                       7-[[2-[4-(pyridiniummethyl)-                                 phenyl]-2-sulfoacetyl]-                                                                        phenyl]-2-sulfoacetyl]-                                      amino]-2-[(1,3,4-thia-                                                                         amino]-3-[(1,3,4-thiadia-                                    diazol-5-ylthio)methyl]-                                                                       zol-5-ylthio)methyl]-8-oxo-                                  8-oxo-5-thia-1-azabicy-                                                                        5-thia-1-azabicyclo[4.2.0]-                                  clo[4.2.0]oct-2-ene-2-                                                                         oct-2-ene-2-carboxylic acid                                  carboxylic acid                                                               7-[[4-[4-(chloromethyl)-                                                                       7-[[4-[4-(pyridiniummethyl)-                                 phenyl]-2-sulfobutyryl]-                                                                       2-sulfobutyryl]amino]-3-[(3-                                 amino]-3-[(3-methyl-1,2,-                                                                      methyl-1,2,4-thiadiazol-5-yl-                                4-thiadiazol-5-ylthio)-                                                                        thio)methyl]  -8-oxo-5-thia-1-                               methyl]-8-oxo-5-thia-1-                                                                        azabicyclo[4.2.0]oct-2-ene-                                  azabicyclo[4.2.0]oct-2-                                                                        2-carboxylic acid                                            ene-2-carboxylic acid                                                         7-[[4-[4-(chloromethyl)-                                                                       7-[[4-[4-(pyridiniummethyl)-                                 phenoxy]-2-carboxybu-                                                                          phenoxyl]-2-carboxybutyryl]-                                 tyryl]amino]-3-[(tetra-                                                                        amino]-3-[(tetrazol-5-ylthio)-                               zol-5-ylthio)methyl]-8-                                                                        methyl]-8-oxo-5-thia-1-azabi-                                oxo-5-thia-1-azabicyclo-                                                                       cyclo[4.2.0]oct-2-ene-2-car-                                 [4.2.0]oct-2-ene-2-car-                                                                        boxylic acid                                                 boxylic acid                                                                  7-[[3-[4-(chloromethyl)-                                                                       7-[[3-[4-(pyridiniummethyl)-                                 anilno]-2-caboxypro-                                                                           anilino]-2-carboxypropionyl]-                                pionyl]amino]-3-[1-                                                                            amino]-3-[(1-methyltetrazol-                                 methyltetrazol-5-yltio)-                                                                       5-ylthio)methyl]-8-oxo-5-                                    methyl]-8-oxo-5-thia-1-                                                                        thia-1-azabicyclo[4.2.0]oct-                                 azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         7-[[3-[5-(chloromethyl)-                                                                       7-[[3-[5-(pyridiniummethyl)-                                 2-thienyl]-2-carboxypro-                                                                       2-thienyl]-2-carboxypropionyl]-                              pionyl]amino]-3-[(2-                                                                           amino]3-3[(2-methyl-1,3,4-oxa-                               methyl-1,3,4-oxadiazol-                                                                        diazol-5-ylthio)methyl]-8-oxo-                               5-ylthio)methyl]-8-oxo-                                                                        5-thia-1-azabicyclo[4.2.0]-                                  5-thia-1-azabicyclo-                                                                           oct-2-ene-2-carboxylic acid                                  [4.2.0]oct-2-ene-2-car-                                                       boxylic acid                                                                  3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-carboxyacetyl]-                                                                      2-carboxyacetyl]amino]-8-oxo-                                amino-8-oxo-5-thia-1-aza-                                                                      5-thia-1-azabicyclo[4.2.0]oct-                               bicyclo[4.2.0]oct-2-ene-                                                                       2-ene-2-carboxylic acid pro-                                 2-carboxylic acid propi-                                                                       pionyloxymethyl ester                                        onyloxymethyl ester                                                           7-[[2-[4-(chloromethyl)-                                                                       7-[[2-[4-(pyridiniummethyl)-                                 phenyl]-2-carboxyacetyl]-                                                                      phenyl]-2-carboxyacetyl]amino]-                              amino]-3-[(2-methyl-1,3,4-                                                                     3-[(2-methyl-1,3,4-thiadiazol-                               thiadiazol-5-ylthio)-                                                                          5-ylthio)methyl]-8-oxo-5-thia-                               methyl] -8-oxo-5-thia-1-                                                                       1-azabicyclo[4.2.0]oct-2-ene-2-                              azabicyclo[4.2.0]oct-2-                                                                        carboxylic acid                                              ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl)-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]acetyl]amino]-7-                                                                        acetyl]amino]-7-methoxy-8-oxo-                               methoxy-8-oxo-5-thia-1-                                                                        5-thia-1-azabicyclo[4.2.0]oct-                               azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         ______________________________________                                    

EXAMPLE 273-[(Acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

(A) A mixture of 7.5 g of3-[(acetyloxy)methyl]-7-[[2-[4-(chloromethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and 200 ml of pyridine is stirred at room temperature for 24 hoursafter which the solvent is removed in vacuo at room temperature. Theproduct is trituratedwith 2:1 benzene-acetone to give the aminepyridiniummethyl derivative.

(B) At 0° C. 10 ml of trifluoroacetic acid is added to 5 g of3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid wherein the2-amino group is protected with tert-butoxycarbonylobtained in (A) above. The mixture is stirred for several minutes undera nitrogen atmosphere at room temperature. The excess trifluoroaceticacid is removed in vacuo and the remaining residue is triturated withdiethylether then dissolved in 175 ml of water. The solution is filteredand the pH of the filtrate adjusted to 5.5 by adding Amberlite IR 4Bresin that has been washed several times with water. The resin isfiltered off and the water concentrated in vacuo. A precipitate formsfrom the concentrate which is removed and washed with ethanol to give3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

In a similar manner when appropriate amounts of chloromethylatedderivatives listed in the following Table XII are reacted with pyridineasin Example 28(A) and the resulting product subsequently treated as inExample 28(B), the respective pyridiniummethyl substituted cephalosporinproducts listed in Table XII are obtained.

                  TABLE XII                                                       ______________________________________                                        CHLOROMETHYL SUBSTI-                                                                           PYRIDINIUMMETHYL SUB-                                        TUTED CEPHALOSPORIN                                                                            STITUTED CEPHALOSPORIN                                       DERIVATIVE       DERIVATIVE                                                   ______________________________________                                        3-[(acetyloxy)methyl]-7-[[3-                                                                   3-[(acetyloxy)methyl-7-[[3-                                  [4-(chloromethyl)phenyl]-2-                                                                    [4-(pyridiniummethyl)phenyl]-                                aminopropionyl]amino]-8-oxo-                                                                   2-aminopropionyl]amino]-8-                                   5-thia-1-azabicyclo[4.2.0]-                                                                    oxo-5-thia-1-azabicyclo-                                     oct-2-ene-2-carboxylic acid                                                                    [4.2.0]oct-2-ene-2-carboxylic                                                 acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]-                                phenyl]-2-aminobutyryl]-                                                                       2-aminobutyryl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]oct-                               azabicyclo[4.2.0 oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-aminobutyryl]-                                                                      2-aminobutyryl]amino]-8-oxo-                                 amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[acetyloxy)methyl]-7-[[3-                                  [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenoxy]-                               phenoxy]-2-aminopropi-                                                                         2-aminopropionyl]amino]-8-oxo-                               onyl]amino]-8-oxo-5-thia-                                                                      5-thia-1-azabicyclo[4.2.0]-                                  1-azabicyclo[4.2.0]oct-                                                                        oct-2-ene-2-carboxylic acid                                  2-ene-2-carboxylic acid                                                       3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)anilino]-                               anilino]-2-aminobutyryl]-                                                                      2-aminobutyryl]-8-oxo-5-thia-                                8-oxo-5-thia-1-azabi-                                                                          1-azabicyclo[4.2.0]oct-2-ene-                                cyclo[4.2.0]oct-2-ene-                                                                         2-carboxylic acid                                            2-carboxylic acid                                                             3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[4-(Chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]-2-aminobu-                                                                         thio]-2-aminobutyryl]amino]  -                               tyryl]amino]-8-oxo-5-                                                                          8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[3-                                 [[3-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl-                                 phenylthio]-2-aminopro-                                                                        thio]-2-aminopropionyl]amino]-                               pionyl]amino]-8-oxo-5-                                                                         8-oxo-5-thia-1-azabicyclo-                                   thia-1-azabicyclo[4.2.0]-                                                                      [4.2.0]oct-2-ene-2-carboxylic                                oct-2-ene-2-carboxylic                                                                         acid                                                         acid                                                                          3-[acetyloxy)methyl]-7-                                                                        3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thi-                                 thienyl]-2-aminoacetyl]-                                                                       enyl]-2-aminoacetyl]amino]-8-                                amino]-8-oxo-5-thia-1-                                                                         oxo-5-thia-1-azabicyclo[4.2.0]-                              azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[ [ 3-                               [[3-[5-(chloromethyl)-2-                                                                       [5-(pyridiniummethyl)-2-thio                                 thienyl]-2-aminopropionyl]-                                                                    enyl]-2-aminopropionyl]amino]-                               amino]-8-oxo-5-thia-1-aza-                                                                     8-oxo-5-thia-1-azabicyclo-                                   bicyclo[4.2.0]oct-2-ene-2-                                                                     [4.2.0]oct-2-ene-2-carboxylic                                carboxylic acid  acid                                                         3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[4-                                 [[4-[5-(chloromethyl)-2-                                                                       [5-(pyridiniumethyl)-2-thi-                                  thienyl]-2-aminobutyryl]-                                                                      enyl]-2-aminobutyryl]amino]-                                 amino]-8-oxo-5-thia-1-                                                                         8-oxo-5-thia-1-azabicyclo-                                   azabicyclo[4.2.0]oct-2-                                                                        [4.2.0]oct-2-ene-2-carboxlic                                 ene-2-carboxylic acid                                                                          acid                                                         3-[(2-methyl-1,3,4-thia-                                                                       3-[(2-methyl-1,3,4-thiadiazol-                               diazol-5-ylthio)methyl]-7-                                                                     5-ylthio)methyl-7-[[2-[4-                                    [[3- [4-(chloromethyl)-                                                                        (pyridiniummethyl)phenyl]-2-                                 phenyl]-2-aminoacetyl]-                                                                        aminoacetyl]amino]-8-oxo-5-                                  amino]-oxo-5-thia-1-                                                                           thia-1-azabicyclo[4.2.0]oct-                                 azabicyclo[4.2.0]oct-2-                                                                        2-ene-2-carboxylic acid                                      ene-2-carboxylic acid                                                         3-[(1,3,4-thiadiazol-                                                                          3-[(1,3,4-thiadiazol-                                        5-ylthio)methyl]-7-[[3-                                                                        5-ylthio)methyl]-7-[[3-                                      [4-(chloromethyl)phenyl]-                                                                      [4-(pyridiniummethyl)phenyl]-                                2-aminopropionyl]amino]-                                                                       2-aminopropionyl]amino]-                                     8-oxo-5-thia-1-azabicy-                                                                        8-oxo-5-thia-1-azabicyclo-                                   cyclo[4.2.0]oct-2-ene-2-                                                                       [4.2.0]oct-2-ene-2-carboxylic                                carboxylic acid  acid                                                         3-[(3-methyl-1,2,4-thia-                                                                       3-[(3-methyl-1,2,4-thiadia-                                  diazol-5-ylthio)methyl]-7-                                                                     zol-5-ylthio)methyl]-7- [4-                                  [[4-[4-(chloromethyl)phen-                                                                     [4-(pyridiniummethyl)phenoxy]-                               oxy]-2-aminobutyryl]amino]-                                                                    2-aminobutyryl]amino]-8-oxo-                                 8-oxo-5-thia-1-azabicy-                                                                        5-thia-1-azabicyclo[4.2.0]-                                  clo[4.2.0]oct-2-ene-2-car-                                                                     oct-2-ene-2-carboxylic acid                                  boxylic acid                                                                  3-[(tetrazol-5-ylthio)-                                                                        3-[(tetrazol-5-ylthio)-                                      methyl]-7-[[3-[4-(chloro-                                                                      methyl]-7-[[3-[4-(pyridinium-                                methyl]phenoxy]-2-amino-                                                                       methyl]phenoxy]-2-amino-pro-                                 propionyl]amino]-8-oxo-5-                                                                      pionyl]amino]-8-oxo-5-thia-                                  thia-1-azabicyclo[4.2.0]-                                                                      1-azabicyclo[4.2.0]oct-2-                                    oct-2-ene-2-carboxylic                                                                         ene-2-carboxylic acid                                        acid                                                                          3-[(1-methyltetrazol-5-                                                                        3-[(1-methyltetrazol-5-yl-                                   ylthio)methyl]-7-[[4-[4-                                                                       thio)methyl]-7-[[4-[4-(pyri-                                 (chloromethyl)phenylthio]-                                                                     diniummethyl)phenylthio]-2-                                  2-aminobutyryl]amino]-8-                                                                       aminobutyryl]amino]-8-oxo-                                   oxo-5-thia-1-azabicyclo-                                                                       5-thia-1-azabicyclo[4.2.0]-                                  [4.2.0]oct-2-ene-2-car-                                                                        oct-2-ene-2-carboxylic acid                                  boxylic acid                                                                  3-[(2-methyl-1,3,4-oxa-                                                                        3-[(2-methyl-1,3,4-oxadia-                                   diazol-5-ylthiomethyl]-                                                                        zol-5-ylthio)methyl]-7-[[2-                                  7-[[2-[5-(chloromethyl)-2-                                                                     [5-(pyridiniummethyl)-2-                                     thienyl]-2-aminoacetyl]-                                                                       thienyl]-2-aminoacetyl]-                                     amino]-8-oxo-5-thia-1-                                                                         amino]-8-oxo-5-thia-1-aza-                                   azabicyclo[4.2.0]oct-2-                                                                        bityclo[4.2.0]oct-2-ene-2-                                   ene-2-carboxylic acid                                                                          carboxylic acid                                              3-[(acetyloxy)methyl]-7-                                                                       3-[(acetyloxy)methyl]-7-[[2-                                 [[2-[4-(chloromethyl)-                                                                         [4-(pyridiniummethyl)phenyl]  -                              phenyl]2-aminoacetyl]-                                                                         2-aminoacetyl]amino]-8-oxo-                                  amino]-8-oxo-5-thia-1-                                                                         5-thia-1-azabicyclo[4.2.0]-                                  azabicyclo[4.2.0]oct-2-                                                                        oct-2-ene-2-carboxylic acid                                  ene-2-carboxylic acid                                                                          pivalyloxymethyl ester                                       pivalyloxymethyl ester                                                        3-methyl-7-[[2-[4-(chloro-                                                                     3-methyl-7-[[2-[4-(pyridin-                                  methyl)phenyl]-2-aminoace-                                                                     iummethyl)phenyl]-2-amino-                                   tyl]amino]-8-oxo-5-thia-1-                                                                     acetyl]amino]-8-oxo-5-thia-                                  azabicyclo[4.2.0]oct-2-ene-                                                                    1-azabicyclo[4.2.0]oct-2-                                    2-carboxylic acid                                                                              ene-2-carboxylic acid                                        3-methyl-7-[[2-[4-(chloro-                                                                     3-methyl-7-[[2-[4-(pyridin-                                  methyl)phenyl]-2-aminoace-                                                                     iummethyl)phenyl]-2-amino-                                   tyl]amino]-8-oxo-5-thia-1-                                                                     acetyl]amino] -8-oxo-5-thia-                                 azabicyclo[4.2.0]oct-2-ene-                                                                    1-azabicyclo[4.2.0]oct-2-                                    2-carboxylic acid pivalyl-                                                                     ene-2-carboxylic acid piv-                                   oxymethyl ester  alyloxymethyl ester                                          ______________________________________                                    

EXAMPLE 283-[(Acetyloxy)methyl]-7-[[2-[4(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-ethoxycarbonyl-N-methylaminomethyl ester

A mixture of 1.2 g of the inner salt of3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and 0.5 g of N-chloromethyl-N-methylurethane in 40 ml ofdimethylformamide is stirred at room temperature for 2 hours. Themixture is poured into ice-water and decanted. The aqueous layer iswashed with 75 ml of ethyl acetate filteredand evaporated. The residueis triturated with ether to give3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid N-ethoxycarbonyl-N-methylaminomethyl ester.

EXAMPLE 293-[(Acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-pivalyloxybenzyl ester

To a solution of 1.8 g of3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid hydrochloride in 25ml of dimethyl formamide is added 0.78 g ofp-pivalyloxybenzyl alcohol followed by cooling to 0° C. after which 3.7mole of dicyclohexylcarbodiimide in 7.5 ml of dimethyl formamide isadded dropwisewith stirring. The reaction mixture is stirred for 1 hourat 0° C. and for an additional 4 hours at room temperature. The formeddicyclohexylurea is removed by filtration. The filtrate is evaporated invacuo to give an oil which is triturated with ether to give3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid p-pivalyloxybenzyl ester.

EXAMPLE 303-[(Acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-(5-indanyloxycarbonyl)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

To 25.3 m mole of3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in 35 ml dioxane is added 6N hydrochloric acid to give a pH of 2.5.Then 24.1 molesN,N'-dicyclohexylcarbodiimide in 35 ml dioxane is addedand the mixture is stirred at room temperature for 15 to 20 minutesfollowed by the addition of 24.1 m moles of 5-indanol. The mixture isstirred for 4 hours. The formed N,N'-dicyclohexylurea is removed byfiltration and the filtrate is extracted three times with methylisobutyl ketone. The organic extract is washed with water, dried overmagnesium sulfate and concentrated to dryness in vacuo to yield3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-(5-indanyloxycarbonyl)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

EXAMPLE 31 p-Pyridiniummethylphenylacetic acid hydrochloride

A solution of 0.25 g (1.33 mM) of p-chloromethylphenylacetic acid and 10mlof pyridine is refluxed in 65 ml of ethanol for 31/2 hours after whichthe solvent is removed in vacuo at room temperature. The resulting oilis triturated with 110 ml of acetone/benzene (1:2) to givep-pyridiniummethylphenylacetic acid hydrochloride.

In a similar manner, when an appropriate amount of a chloromethylderivative listed in Table I, Table II and Table III is substituted forp-chlorophenylacetic acid in the above procedure, the correspondingpyridiniummethyl acid derivative is obtained.

EXAMPLE 323-[(Acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid

A mixture of 1.2 grams of3-[(acetyloxy)methyl]-7-amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and 1.2 grams of p-pyridiniummethylphenylacetic acid chloride HClin 55 ml of dimethylformamide containing 0.5 ml of triethylamine isstirred for 11/2 hours after which the solvent is removed yielding3-[(acetyloxy)methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid which can be further purified by trituration with chloroform.

In a similar manner as in the above procedure when an appropriate amountofa pyridiniummethyl acid derivative described in Example 31 issubstituted for p-pyridiniummethylphenylacetic acid the correspondingpyridiniummethylsubstituted cephalosporin derivative is obtained.

EXAMPLE 333-(2-methyl-1,3,4-thiadiazol-5-ylthio)-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]-oct-2-ene-2-carboxylicacid

A solution of 3 mM of the inner salt of3-[(acetyloxymethyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid in 100 ml of water is treated with 3 mM of sodium bicarbonate and 6mM of 2-methyl-1,3,4-thiadiazol-5-yl-thio at 70° C. under nitrogen for31/2 hours. The water is removed in vacuo and the residue is trituratedwith ether then filtered and dried in a vacuum desiccator to give3-(2-methyl-1,3,4-thiadiazol-5-ylthio)-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid.

In a similar manner other compounds of this invention wherein the3-position of the cephalosporin molecule is substituted with aheterocyclicthiomethyl group as described in Formula I can be preparedby solvolysis of the corresponding 3-acetoxy derivative with anappropriate heterocyclicthiol compound.

We claim:
 1. A compound selected from a base of the formula: ##STR20##wherein Aryl is selected from phenyl and 2-thienyl; Y is selected fromhydrogen, chlorine, bromine, a straight or branched lower alkyl group offrom 1 to 4 carbon atoms, and a lower alkoxy group of from 1 to 4 carbonatoms with the proviso that when Aryl is 2-thienyl, Y is hydrogen; Z isselected from a bond, oxygen, sulfur and imino with the proviso thatwhen Aryl is 2-thienyl, Z is a bond; W is selected from hydrogen,methyl, amino, hydroxy, SO₃ H and COOR₁ wherein R₁ is selected fromhydrogen and 5-indanyl; n is zero, 1 or 2 with the proviso that when Wis other than hydrogen or methyl, and Z is other than a bond, n is notzero; R₂ is selected from hydrogen or methoxy and is either cis- ortrans-; M is selected from an anion; hydrogen alkanoyloxymethyl whereinthe alkanoyl moiety contains from 1 to 5 carbon atoms and may bestraight or branched; alkanoylaminomethyl wherein the alkanoyl moietycontains from 1 to 5 carbon atoms and may be straight or branched andwherein the amino nitrogen may be substituted with an alkyl group offrom 1 to 4 carbon atoms; alkoxycarbonylaminomethyl wherein the alkoxymoiety contains from 1 to 4 carbon atoms and may be straight or branchedand wherein the amino nitrogen atom may be substituted with an alkylgroup of from 1 to 4 carbon atoms; p-(alkanoyloxy)benzyl wherein thealkanoyl moiety contains from 1 to 5 carbon atoms and may be straight orbranched; or aminoalkanoyloxymethyl wherein the alkanoyl moiety containsfrom 2 to 15 carbon atoms, and the amino nitrogen atom may be mono- ordi-substituted with a lower alkyl group of from 1 to 4 carbon atoms;with the proviso that when M is other than an anion the compound existsas a salt of a pharmaceutically acceptable inorganic or organic acid; Xis hydrogen or acetoxy; and pharmaceutically acceptable salts thereof.2. A compound of claim 1 wherein Aryl is phenyl.
 3. A compound of claim2 wherein the R₂ substituent is in the cis-position.
 4. A compound ofclaim 1 wherein Aryl is 2-thienyl.
 5. A compound of claim 4 wherein theR₂ substituent is in the cis-position.
 6. A compound of claim 1 whereinW is hydrogen.
 7. A compound of claim 6 wherein Z is a bond.
 8. Acompound of claim 6 wherein Z is oxygen or sulfur.
 9. A compound ofclaim 6 wherein Z is imino.
 10. A compound of claim 1 wherein W ismethyl.
 11. A compound of claim 10 wherein Z is a bond.
 12. A compoundof claim 10 wherein Z is oxygen or sulfur.
 13. A compound of claim 10wherein Z is imino.
 14. A compound of claim 1 wherein W is hydroxy. 15.A compound of claim 14 wherein Z is a bond.
 16. A compound of claim 14wherein Z is oxygen or sulfur.
 17. A compound of claim 14 wherein Z isimino.
 18. A compound of claim 1 wherein W is amino.
 19. A compound ofclaim 18 wherein Z is a bond.
 20. A compound of claim 18 wherein Z isoxygen or sulfur.
 21. A compound of claim 18 wherein Z is imino.
 22. Acompound of claim 1 wherein W is COOR₁ or SO₃ H.
 23. A compound of claim22 wherein Z is a bond.
 24. A compound of claim 22 wherein Z is oxygenor sulfur.
 25. A compound of claim 22 wherein Z is imino.
 26. A compoundselected from a base of the formula: ##STR21## wherein W' is selectedfrom hydrogen, hydroxy, amino, COOH, or SO₃ H; X' is hydrogen oracetoxy; and wherein the hydrogen atoms at the 6- and 7- positions arecis to one another; and pharmaceutically acceptable salts thereof.
 27. Acompound of claim 26 which is3-[(acetyloxy)-methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and pharmaceutically acceptable salts thereof.
 28. A compound ofclaim 26 which is3-[(acetyloxy)-methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-hydroxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and pharmaceutically acceptable salts thereof.
 29. A compound ofclaim 26 which is3-[(acetyloxy)-methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-aminoacetyl]-amino]-8-oxo-5-thia-1azabicyclo[[4.2.0]oct-2-ene-2-carboxylicacid and pharmaceutically acceptable salts thereof.
 30. A compound ofclaim 26 which is3-[(acetyloxy)-methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-carboxyacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxlicacid and pharmaceutically acceptable salts thereof.
 31. A compound ofclaim 26 which is3-[(acetyloxy)-methyl]-7-[[2-[4-(pyridiniummethyl)phenyl]-2-sulfoacetyl]-amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid and pharmaceutically acceptable salts thereof.
 32. A compound ofclaim 26 which is3-methyl-7-[[2-[4-(pyridiniummethyl)phenyl]-2-aminoacetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid and pharmaceutically acceptable saltsthereof.